LOCUS       V036571                 9196 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036571
VERSION     V036571
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..4050
                     /label="Alcam(NM_009655)"
                     /note="Alcam(NM_009655)"
                     /gene="Alcam"
     CDS             4057..4122
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4166..4377
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4390..4658
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4685..5284
                     /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    5291..5879
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5953..6186
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6258..6379
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6398..6533
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6571..6587)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6595..6611
                     /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(6619..6649)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6664..6685
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6973..7561)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7732..8592)
                     /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(8593..8697)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9171..9187
                     /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 catggcatct aaggtgtccc cttcttgccg tctggttttc
     2341 tgcctcttga tctccgccgc ggtcctcaga ccaggccttg gatggtacac tgtcaactca
     2401 gcatatggag acaccattgt catgccttgc cgtctcgatg tacctcagaa cctcatgttt
     2461 ggcaaatgga aatatgaaaa gcccgatggg tccccagtat ttattgcatt ccgatcttct
     2521 acgaagaaaa gtgtgcagta tgatgatgtc ccagagtaca aggacagact gagcctctca
     2581 gaaaactaca ctttgtctat cgccaatgct aagatcagtg atgagaagag atttgtgtgc
     2641 atgctagtga ccgaggacaa tgtgtttgaa gcacctaccc tagtcaaggt gttcaagcaa
     2701 ccatctaaac ctgaaattgt aaacaaagca ccctttcttg aaacagatca gctgaaaaag
     2761 ttaggtgact gcatttcaag agacagttac ccagatggca acatcacatg gtacaggaat
     2821 ggcaaagtgc tgcaacctgt ggaaggagag gtggccatac tttttaaaaa ggaaattgat
     2881 ccaggaactc agttgtatac cgtgacttcc tccttggagt acaagacaac cagatctgac
     2941 atacaaatgc cattcacctg ttctgtgaca tattatggac cctccggcca gaaaacaatt
     3001 tattccgaac aagaaatctt tgatatttac tatcctacag agcaggtgac aatacaagtg
     3061 ctgccaccaa aaaatgccat caaagaaggg gacaacatca cccttcagtg tttggggaat
     3121 ggcaacccac ctcctgagga gttcatgttt tacttaccag gacagcctga aggcataaga
     3181 agctcaaaca cttacacact gacagatgtg agacgcaatg ccacagggga ctacaaatgc
     3241 tcactcattg acaaaaggaa catggcggct tcaacgacca tcacagttca ctacttggat
     3301 ttatccttaa acccaagtgg ggaggtgacc aagcagattg gcgataccct gcctgtgtca
     3361 tgcacaatat ctgcaagtag gaacgcgact gtggtgtgga tgaaggataa catccggctc
     3421 cggtccagcc cttccttttc cagtcttcat tatcaggatg ctgggaacta tgtctgtgaa
     3481 actgctcttc aggaggttga gggactgaag aaaagggagt cgctgaccct cattgtagaa
     3541 gggaaacctc aaatcaaaat gactaagaaa actgacccca gtggactgtc taaaactata
     3601 atctgccacg tggaagggtt tccaaagcca gctatacatt ggaccattac cggcagtggg
     3661 agcgtcataa accaaacaga ggagtctcct tatattaatg gcaggtatta tagtaaaatt
     3721 atcatttccc ctgaggagaa tgtaacatta acttgcacag cagaaaacca actggagaga
     3781 acagtaaact ccctgaatgt ctctgcgata agtattccag agcacgatga ggcagacgat
     3841 ataagtgatg aaaacagaga aaaggtgaat gaccaggcaa aactgattgt gggaattgtc
     3901 gttggtctcc tcctcgctgc cctcgttgct ggtgtcgtct actggctgta catgaagaag
     3961 tccaaaactg catcaaaaca tgtaaacaag gaccttggta atatggagga aaataaaaag
     4021 ctggaagaaa acaatcacaa aacagaagcc ggatccgact acaaagacca tgacggtgat
     4081 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc
     4141 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4201 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     4261 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4321 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     4381 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     4441 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     4501 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     4561 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     4621 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     4681 ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     4741 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     4801 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4861 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4921 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4981 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5041 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5101 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5161 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5221 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5281 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5341 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5401 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5461 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5521 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5581 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5641 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     5701 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5761 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5821 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5881 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5941 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt
     6001 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6061 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6121 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6181 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     6241 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6301 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6361 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     6421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6481 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     6541 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6601 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6661 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6721 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6781 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7381 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7861 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8041 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8341 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8701 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     8761 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     8821 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     8881 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     8941 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     9001 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     9061 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9121 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     9181 ggccagtgcc aagctg
//
