LOCUS       V032398                 9248 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032398
VERSION     V032398
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..3249
                     /label="Spp1(NM_001204201)"
                     /note="Spp1(NM_001204201)"
                     /gene="Spp1"
     CDS             3256..3282
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3286..3312
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3316..3342
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3407..3618
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3631..3899
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3928..3937
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3958..4620
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4690..4743
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4744..5343
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5344..5931
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6005..6238
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6310..6431
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6450..6585
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6623..6639)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6647..6663
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6671..6701)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6716..6737
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7025..7613)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7784..8644)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8645..8749)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9223..9239
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatga ggctgcagtt ctcctggctg
     2341 aattctgagg gactaactac gaccatgaga ttggcagtga tttgcttttg cctgtttggc
     2401 attgcctcct ccctcccggt gaaagtgact gattctggca gctcagagga gaagaagctt
     2461 tacagcctgc acccagatcc tatagccaca tggctggtgc ctgacccatc tcagaagcag
     2521 aatctccttg cgccacagaa tgctgtgtcc tctgaagaaa aggatgactt taagcaagaa
     2581 actcttccaa gcaattccaa tgaaagccat gaccacatgg acgacgatga tgacgatgat
     2641 gatgacgatg gagaccatgc agagagcgag gattctgtgg actcggatga atctgacgaa
     2701 tctcaccatt cggatgagtc tgatgagacc gtcactgcta gtacacaagc agacactttc
     2761 actccaatcg tccctacagt cgatgtcccc aacggccgag gtgatagctt ggcttatgga
     2821 ctgaggtcaa agtctaggag tttccaggtt tctgatgaac agtatcctga tgccacagat
     2881 gaggacctca cctctcacat gaagagcggt gagtctaagg agtccctcga tgtcatccct
     2941 gttgcccagc ttctgagcat gccctctgat caggacaaca acggaaaggg cagccatgag
     3001 tcaagtcagc tggatgaacc aagtctggaa acacacagac ttgagcattc caaagagagc
     3061 caggagagtg ccgatcagtc ggatgtgatc gatagtcaag caagttccaa agccagcctg
     3121 gaacatcaga gccacaagtt tcacagccac aaggacaagc tagtcctaga ccctaagagt
     3181 aaggaagatg ataggtatct gaaattccga atttctcatg aattagagag ttcatcttct
     3241 gaggtcaacc tcgagtaccc atacgacgta cctgattatg caacttatcc ttatgatgta
     3301 cctgattatg ctagctatcc ttatgatgtc ccagactacg catagggatc cctgcgatat
     3361 ccagcgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     3421 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     3481 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3541 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     3601 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     3661 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     3721 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     3781 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     3841 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     3901 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc
     3961 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg
     4021 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa
     4081 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac
     4141 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc
     4201 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc
     4261 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc
     4321 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg
     4381 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac
     4441 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc
     4501 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc
     4561 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc
     4621 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc
     4681 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     4741 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     4801 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     4861 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     4921 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     4981 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5041 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     5101 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5161 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     5221 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5281 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5341 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5401 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5461 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5521 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5581 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5641 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5701 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     5761 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5821 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5881 ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct
     5941 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     6001 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg
     6061 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6121 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6181 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6241 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6301 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6361 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6421 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc
     6481 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6541 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag
     6601 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6661 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6721 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6781 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6841 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6901 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6961 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7021 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7081 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7141 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7201 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7261 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7321 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7381 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7441 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7501 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7561 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7621 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7681 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7741 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7801 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7861 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7921 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7981 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8041 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8101 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8161 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8221 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8281 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8341 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8401 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8461 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8521 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8581 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8641 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8701 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8761 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     8821 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     8881 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     8941 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     9001 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     9061 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     9121 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     9181 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     9241 ccaagctg
//
