LOCUS       V046289                 9378 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046289
VERSION     V046289
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3397
                     /label="Ccn4(NM_018865)"
                     /note="Ccn4(NM_018865)"
                     /gene="Ccn4"
     CDS             3404..3493
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3537..3748
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3761..4029
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4058..4067
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4088..4750
                     /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             4820..4873
                     /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             4874..5473
                     /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    5474..6061
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6135..6368
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6440..6561
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6580..6715
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6753..6769)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6777..6793
                     /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(6801..6831)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6846..6867
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7155..7743)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7914..8774)
                     /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(8775..8879)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9353..9369
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatga ggtggctcct gccctggacg ctggcagccg tggcagtcct
     2341 gagggtgggc aacatcctgg ccacggccct ctctccaacc cccacaacaa tgaccttcac
     2401 cccagcacca ctagaggaaa cgactacacg ccccgaattc tgcaagtggc catgtgagtg
     2461 cccacaatcc ccacctcgct gcccactggg cgtcagccta atcacagatg gctgtgaatg
     2521 ctgtaagata tgtgcccagc agcttgggga caactgcaca gaggctgcca tctgtgaccc
     2581 acaccggggc ctctactgcg attacagtgg ggatcgcccg aggtacgcaa taggagtgtg
     2641 tgcacaggtg gtcggtgtgg gctgtgtcct ggatggcgta cgctacacca atggcgagtc
     2701 cttccaaccc aactgcaggt acaactgtac ctgcattgat ggcacggtgg gctgcacacc
     2761 gctgtgccta agccccaggc ccccacgcct ctggtgccgc cagccccggc acgtgagagt
     2821 ccctggccag tgctgtgagc agtgggtgtg tgatgatgac gcaaggagac cacgccagac
     2881 tgcactgttg gacaccagag cctttgcagc gtcaggcgcc gtggagcaac ggtatgagaa
     2941 ctgcatagcc tacactagtc cctggagccc ctgctctacc acctgtggcc taggtatctc
     3001 cactcggatc tctaacgtca atgcccggtg ctggccagag caggaaagtc gcctctgcaa
     3061 cctgcggcca tgtgatgtgg acatccaact acacatcaag gcagggaaga aatgcctggc
     3121 tgtgtaccag ccagaggagg ccacgaactt cactctcgca ggctgtgtca gcacacgcac
     3181 ctaccgaccc aagtactgcg gagtctgtac tgacaatagg tgttgcatcc cctacaagtc
     3241 caagaccatc agtgtggatt tccagtgtcc agaggggcca ggtttctccc ggcaggtcct
     3301 atggattaat gcttgcttct gcaacctgag ctgcaggaat cctaacgata tctttgctga
     3361 cttggaatct taccctgact tcgaagagat tgccaatgga tccgaacaaa aactaatatc
     3421 agaggaagat ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc
     3481 cgaggaggac ttatagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3541 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3601 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     3661 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     3721 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     3781 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     3841 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     3901 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     3961 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     4021 ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag
     4081 cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt
     4141 cgagctggtg ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa
     4201 gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg
     4261 cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa
     4321 caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt
     4381 gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac
     4441 cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt
     4501 ggagcacctg caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag
     4561 cctgcgcgac ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc
     4621 catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga
     4681 gctgcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat
     4741 cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc
     4801 cggctccacc ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga
     4861 gaatcccggc cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     4921 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     4981 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     5041 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     5101 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     5161 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     5221 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     5281 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     5341 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     5401 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     5461 gcccggtgcc tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5521 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5581 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5641 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5701 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5761 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5821 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     5881 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5941 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6001 tcttccgcgt ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg
     6061 cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6121 agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc
     6181 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6241 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6301 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6361 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6421 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     6481 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6541 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     6601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6661 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac
     6721 ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6781 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6841 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6901 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6961 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7021 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7081 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7141 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7201 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7261 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7321 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7381 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7441 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7501 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7561 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7621 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7681 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7741 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7801 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7861 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7921 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7981 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8041 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8101 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8161 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8221 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8281 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8341 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8401 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8461 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8521 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8581 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8641 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8701 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8761 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8821 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8881 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     8941 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     9001 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     9061 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     9121 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     9181 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     9241 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     9301 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     9361 acggccagtg ccaagctg
//
