LOCUS       V035242                 8978 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035242
VERSION     V035242
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..3048
                     /label="Klf9(NM_010638)"
                     /note="Klf9(NM_010638)"
                     /gene="Klf9"
     CDS             3049..3072
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        3137..3348
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3361..3629
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3658..3667
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3688..4350
                     /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             4420..4473
                     /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             4474..5073
                     /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    5074..5661
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5735..5968
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6040..6161
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6180..6315
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6353..6369)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6377..6393
                     /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(6401..6431)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6446..6467
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6755..7343)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7514..8374)
                     /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(8375..8479)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8953..8969
                     /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 gccaccatgt ccgcggccgc ctacatggac
     2341 ttcgtggctg cccagtgtct ggtttccatc tccaaccgcg ccgccgtgcc ggagcacggg
     2401 ggcgctccgg aagccgagcg gctgcgacta cctgagcgcg aggtgaccaa ggaacacggt
     2461 gacccggggg acacctggaa ggattattgc acgctggtca ctatcgccaa gagcttgttg
     2521 gacctcaaca aataccgacc catccagacc ccctcggtgt gcagcgacag tctggagagt
     2581 cccgatgagg atataggatc cgacagcgac gtgaccaccg aatctgggtc gagtccttcc
     2641 cacagcccgg aggagagaca ggattctggc agcgcgccca gcccactctc cctcctccac
     2701 tctggagtgg cttcgaaggg gaaacacgcc tccgaaaaga ggcacaagtg cccctacagt
     2761 ggctgtggga aagtctatgg aaaatcctcc catcttaaag cccattacag agtgcataca
     2821 ggtgaacggc cctttccctg cacgtggcca gactgcctta aaaagttctc gcgctcggat
     2881 gagctgaccc gccactaccg gacccacact ggggaaaagc agttccgttg cccactgtgt
     2941 gagaagagat tcatgaggag tgaccatctc accaagcatg cccggcgtca caccgagttc
     3001 catcccagca tgatcaagag atcaaaaaag gctcttgcca gccccttgga ctacaaagac
     3061 gatgacgaca agtgaggatc cctgcgatat ccagcgcggc cgcgaaggat ctgcgatcgc
     3121 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     3181 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     3241 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     3301 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     3361 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     3421 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     3481 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     3541 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     3601 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagacgcc
     3661 accatggaga gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc
     3721 accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc caagcagggc
     3781 cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg
     3841 agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac
     3901 cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag
     3961 gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc
     4021 gacttcaagg tggtgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc
     4081 atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacgt gctggtgggc
     4141 agcttcgccc gcaccttcag cctgcgcgac ggcggctact acagcttcgt ggtggacagc
     4201 cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc
     4261 gccttccgcc gcgtggagga gctgcacagc aacaccgagc tgggcatcgt ggagtaccag
     4321 cacgccttca agacccccat cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc
     4381 gtggacggca ccgccggacc cggctccacc ggatctcgcg agggcagagg aagtcttcta
     4441 acatgcggtg acgtggagga gaatcccggc cctatgaccg agtacaagcc cacggtgcgc
     4501 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4561 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4621 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4681 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4741 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4801 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4861 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4921 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4981 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5041 acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta caaaatttgt
     5101 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5161 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5221 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5281 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5341 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5401 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     5461 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     5521 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     5581 ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc gccctcagac gagtcggatc
     5641 tccctttggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta
     5701 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     5761 aaataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     5821 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5881 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5941 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     6001 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     6061 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6121 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     6181 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6241 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6301 gcttttttgg aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc
     6361 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6421 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6481 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6541 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6601 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6661 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6721 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6781 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6841 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6901 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6961 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7021 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7081 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7141 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7201 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7261 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7321 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7381 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7441 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7501 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7561 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7621 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7681 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7741 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7801 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7861 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7921 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7981 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8041 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8101 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8161 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8221 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8281 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8341 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8401 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8461 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     8521 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     8581 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     8641 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     8701 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     8761 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     8821 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     8881 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     8941 agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
