LOCUS       V032783                 8746 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032783
VERSION     V032783
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..3600
                     /label="CCNB1(NM_031966)"
                     /note="CCNB1(NM_031966)"
                     /gene="CCNB1"
     CDS             3607..3672
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3716..3927
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3940..4208
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4235..4834
                     /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    4841..5429
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5503..5736
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5808..5929
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5948..6083
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6121..6137)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6145..6161
                     /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(6169..6199)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6214..6235
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6523..7111)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7282..8142)
                     /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(8143..8247)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8721..8737
                     /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 catggcgctc cgagtcacca ggaactcgaa aattaatgct
     2341 gaaaataagg cgaagatcaa catggcaggc gcaaagcgcg ttcctacggc ccctgctgca
     2401 acctccaagc ccggactgag gccaagaaca gctcttgggg acattggtaa caaagtcagt
     2461 gaacaactgc aggccaaaat gcctatgaag aaggaagcaa aaccttcagc tactggaaaa
     2521 gtcattgata aaaaactacc aaaacctctt gaaaaggtac ctatgctggt gccagtgcca
     2581 gtgtctgagc cagtgccaga gccagaacct gagccagaac ctgagcctgt taaagaagaa
     2641 aaactttcgc ctgagcctat tttggttgat actgcctctc caagcccaat ggaaacatct
     2701 ggatgtgccc ctgcagaaga agacctgtgt caggctttct ctgatgtaat tcttgcagta
     2761 aatgatgtgg atgcagaaga tggagctgat ccaaaccttt gtagtgaata tgtgaaagat
     2821 atttatgctt atctgagaca acttgaggaa gagcaagcag tcagaccaaa atacctactg
     2881 ggtcgggaag tcactggaaa catgagagcc atcctaattg actggctagt acaggttcaa
     2941 atgaaattca ggttgttgca ggagaccatg tacatgactg tctccattat tgatcggttc
     3001 atgcagaata attgtgtgcc caagaagatg ctgcagctgg ttggtgtcac tgccatgttt
     3061 attgcaagca aatatgaaga aatgtaccct ccagaaattg gtgactttgc ttttgtgact
     3121 gacaacactt atactaagca ccaaatcaga cagatggaaa tgaagattct aagagcttta
     3181 aactttggtc tgggtcggcc tctacctttg cacttccttc ggagagcatc taagattgga
     3241 gaggttgatg tcgagcaaca tactttggcc aaatacctga tggaactaac tatgttggac
     3301 tatgacatgg tgcactttcc tccttctcaa attgcagcag gagctttttg cttagcactg
     3361 aaaattctgg ataatggtga atggacacca actctacaac attacctgtc atatactgaa
     3421 gaatctcttc ttccagttat gcagcacctg gctaagaatg tagtcatggt aaatcaagga
     3481 cttacaaagc acatgactgt caagaacaag tatgccacat cgaagcatgc taagatcagc
     3541 actctaccac agctgaattc tgcactagtt caagatttag ccaaggctgt ggcaaaggtg
     3601 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     3661 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     3721 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     3781 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     3841 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     3901 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     3961 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     4021 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     4081 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     4141 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     4201 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg
     4261 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     4321 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     4381 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4441 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4501 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4561 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4621 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4681 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4741 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4801 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa
     4861 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4921 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4981 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5041 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5101 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5161 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5221 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     5281 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5341 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5401 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     5461 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5521 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5581 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5641 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5701 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     5761 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     5821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     5941 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6001 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6061 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     6121 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6181 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6241 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6301 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6361 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6421 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6481 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6961 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7261 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7321 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7381 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7441 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7501 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7561 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7621 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7681 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7741 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7801 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7861 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7921 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7981 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8041 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8101 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8161 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8221 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     8281 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     8341 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     8401 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     8461 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     8521 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     8581 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     8641 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     8701 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
