LOCUS       V037697                 9519 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037697
VERSION     V037697
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3618
                     /label="TFAP2A(NM_001372066)"
                     /note="TFAP2A(NM_001372066)"
                     /gene="TFAP2A"
     CDS             3625..3690
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3740..3951
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3964..4232
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4267..4959
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
                     /label="TurboRFP"
     CDS             4960..5013
                     /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             5014..5613
                     /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    5614..6202
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6276..6509
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6581..6702
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6721..6856
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6894..6910)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6918..6934
                     /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(6942..6972)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6987..7008
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7296..7884)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8055..8915)
                     /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(8916..9020)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9494..9510
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgaaaatg ctttggaaat tgacggataa tatcaagtac
     2341 gaggactgcg aggaccgtca cgacggcacc agcaacggga cggcacggtt gccccagctg
     2401 ggcactgtag gtcaatctcc ctacacgagc gccccgccgc tgtcccacac ccccaatgcc
     2461 gacttccagc ccccatactt ccccccaccc taccagccta tctaccccca gtcgcaagat
     2521 ccttactccc acgtcaacga cccctacagc ctgaaccccc tgcacgccca gccgcagccg
     2581 cagcacccag gctggcccgg ccagaggcag agccaggagt ctgggctcct gcacacgcac
     2641 cgggggctgc ctcaccagct gtcgggcctg gatcctcgca gggactacag gcggcacgag
     2701 gacctcctgc acggcccaca cgcgctcagc tcaggactcg gagacctctc gatccactcc
     2761 ttacctcacg ccatcgagga ggtcccgcat gtagaagacc cgggtattaa catcccagat
     2821 caaactgtaa ttaagaaagg ccccgtgtcc ctgtccaagt ccaacagcaa tgccgtctcc
     2881 gccatcccta ttaacaagga caacctcttc ggcggcgtgg tgaaccccaa cgaagtcttc
     2941 tgttcagttc cgggtcgcct ctcgctcctc agctccacct cgaagtacaa ggtcacggtg
     3001 gcggaagtgc agcggcggct ctcaccaccc gagtgtctca acgcgtcgct gctgggcgga
     3061 gtgctccgga gggcgaagtc taaaaatgga ggaagatctt taagagaaaa actggacaaa
     3121 ataggattaa atctgcctgc agggagacgt aaagctgcca acgttaccct gctcacatca
     3181 ctagtagagg gagaagctgt ccacctagcc agggactttg ggtacgtgtg cgaaaccgaa
     3241 tttcctgcca aagcagtagc tgaatttctc aaccgacaac attccgatcc caatgagcaa
     3301 gtgacaagaa aaaacatgct cctggctaca aaacagatat gcaaagagtt caccgacctg
     3361 ctggctcagg accgatctcc cctggggaac tcacggccca accccatcct ggagcccggc
     3421 atccagagct gcttgaccca cttcaacctc atctcccacg gcttcggcag ccccgcggtg
     3481 tgtgccgcgg tcacggccct gcagaactat ctcaccgagg ccctcaaggc catggacaaa
     3541 atgtacctca gcaacaaccc caacagccac acggacaaca acgccaaaag cagtgacaaa
     3601 gaggagaagc acagaaagct cgaggactac aaagaccatg acggtgatta taaagatcat
     3661 gacatcgact acaaggatga cgatgacaag tagggatccg cggccgcaag gatctgcgat
     3721 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     3781 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     3841 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     3901 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     3961 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     4021 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     4081 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     4141 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     4201 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac
     4261 gccaccatga gcgagctgat caaggagaac atgcacatga agctgtacat ggagggcacc
     4321 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     4381 cagaccatga agatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     4441 gctaccagct tcatgtacgg cagcaaagcc ttcatcaacc acacccaggg catccccgac
     4501 ttctttaagc agtccttccc tgagggcttc acatgggaga gaatcaccac atacgaagac
     4561 gggggcgtgc tgaccgctac ccaggacacc agcttccaga acggctgcat catctacaac
     4621 gtcaagatca acggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacacgc
     4681 ggctgggagg ccaacaccga gatgctgtac cccgctgacg gcggcctgag aggccacagc
     4741 cagatggccc tgaagctcgt gggcgggggc tacctgcact gctccttcaa gaccacatac
     4801 agatccaaga aacccgctaa gaacctcaag atgcccggct tccacttcgt ggaccacaga
     4861 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga gatggctgtg
     4921 gccaagtact gcgacctccc tagcaaactg gggcacagag agggcagagg aagtcttcta
     4981 acatgcggtg acgtggagga gaatcccggc cctatgaccg agtacaagcc cacggtgcgc
     5041 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     5101 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     5161 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5221 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5281 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5341 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5401 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5461 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5521 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5581 acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta caaaatttgt
     5641 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5701 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5761 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5821 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5881 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5941 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6001 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     6061 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6121 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6181 tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt
     6241 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     6301 aaaataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     6361 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6421 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6481 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     6541 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     6601 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6661 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     6721 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6781 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     6841 ggcttttttg gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag
     6901 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6961 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7021 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7081 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7141 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7201 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7261 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7321 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7381 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7441 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7501 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7561 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7621 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7681 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7741 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7801 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7861 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7921 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7981 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8041 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8101 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8161 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8221 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8281 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8341 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8401 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8461 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8521 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8581 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8641 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8701 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8761 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8821 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8881 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8941 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9001 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     9061 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     9121 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     9181 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     9241 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     9301 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     9361 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     9421 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     9481 cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
