LOCUS       V029705                 9559 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029705
VERSION     V029705
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..4413
                     /label="MARCHF7(NM_001282805)"
                     /note="MARCHF7(NM_001282805)"
                     /gene="MARCHF7"
     CDS             4420..4485
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4529..4740
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4753..5021
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5048..5647
                     /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    5654..6242
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6316..6549
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6621..6742
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6761..6896
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6934..6950)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6958..6974
                     /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(6982..7012)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7027..7048
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7336..7924)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8095..8955)
                     /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(8956..9060)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9534..9550
                     /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 catggagtct aaaccttcaa ggattccaag aagaatttct
     2341 gttcaacctt ccagctcctt aagtgctagg atgatgtctg gaagcagagg aagtagttta
     2401 aatgatacct atcactcaag agactcttca tttagattgg attctgaata tcagtctaca
     2461 tcagcatcag catctgcgtc accatttcaa tctgcatggt atagtgaatc tgagataact
     2521 cagggagcac gctcaagatc gcagaaccag caacgggatc atgattcaaa aagacctaaa
     2581 ctttcctgta caaactgtac tacctcagct gggagaaatg ttggaaatgg tttaaacaca
     2641 ttatcagatt catcttggag gcatagtcaa gttcctagat cttcatcaat ggtacttgga
     2701 tcatttggaa cagacttaat gagagagagg agagatttgg agagaagaac agattcctct
     2761 attagtaatc ttatggatta tagtcaccga agtggtgatt tcacaacttc atcatatgtt
     2821 caagacagag ttccttcata ttcacaagga gcaagaccaa aagaaaactc aatgagcact
     2881 ttacagttga atacatcatc cacaaaccac caattgcctt ctgaacatca gaccatacta
     2941 agttctaggg actccagaaa ttctttaaga tcaaattttt cttcaagaga atcagaatct
     3001 tcccgaagca atacgcagcc tggattttct tacagttcaa gtagagatga agccccaatc
     3061 ataagcaatt cagaaagggt tgtttcatct caaagaccat ttcaagaatc ttctgacaat
     3121 gaaggtaggc ggacaacgag gagattgctg tcacgcatag cttctagcat gtcatctact
     3181 tttttttcac gaagatctag tcaggattcc ttgaatacaa gatcattgaa ttctgaaaat
     3241 tcttacgttt ctccaagaat cttgacagct tcacagtccc gtagtaatgt accatcagct
     3301 tctgaagttc ccgataatag ggcatctgaa gcttctcagg gatttcgatt tcttaggcga
     3361 agatggggtt tgtcatctct tagccacaat catagctctg agtcagattc agaaaatttt
     3421 aaccaagaat ctgaaggtag aaatacagga ccatggttat cttcctcact tagaaataga
     3481 tgcacacctt tgttctctag aaggaggcga gagggaagag atgaatcttc aaggatacct
     3541 acctctgata catcatctag atctcatatt tttagaagag aatcaaatga agtggttcac
     3601 cttgaagcac agaatgatcc tcttggagct gctgccaaca gaccacaagc atctgcagca
     3661 tcaagcagtg ccacaacagg tggctctaca tcagattcgg ctcaaggtgg aagaaataca
     3721 ggaatatcag ggattcttcc tggttcctta ttccggtttg cagtccctcc agcacttggg
     3781 agtaatttga ccgacaatgt catgatcaca gtagatatta ttccttcagg ttggaattca
     3841 gctgatggta aaagtgataa aactaaaagt gcgccttcaa gagatccaga aagattgcag
     3901 aaaataaaag agagcctcct tttagaggac tcagaagaag aagaaggtga cttatgtaga
     3961 atttgtcaaa tggcagctgc atcatcatct aatttgctga tagagccatg caagtgcaca
     4021 ggaagtttgc agtatgtcca ccaagactgt atgaaaaagt ggttacaggc caaaattaac
     4081 tctggttctt cattagaagc tgtaaccacc tgtgaactat gtaaagagaa gttggagctt
     4141 aacctggagg attttgatat tcatgaacta catagagctc atgcaaatga acaagctgag
     4201 tatgagttta tcagctctgg tctctaccta gtggtgttat tgcacttgtg cgaacaaagc
     4261 ttttctgata tgatgggaaa tacaaatgaa ccaagcacac gtgtccgatt tattaacctt
     4321 gcaagaactc ttcaggcaca tatggaagat ctcgaaactt cagaggatga ttccgaagaa
     4381 gacggagacc ataacaggac atttgatatt gccggatccg actacaaaga ccatgacggt
     4441 gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg gccgcgaagg
     4501 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     4561 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac
     4621 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     4681 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     4741 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc
     4801 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg
     4861 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct
     4921 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc
     4981 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct
     5041 acgctagatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     5101 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     5161 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     5221 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     5281 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     5341 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     5401 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     5461 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     5521 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     5581 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     5641 tgcctgagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5701 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5761 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5821 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5881 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5941 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6001 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     6061 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     6121 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6181 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     6241 gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     6301 aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat ctgctttttg
     6361 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     6421 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     6481 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     6541 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     6601 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     6661 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6721 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     6781 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6841 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga
     6901 cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6961 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     7021 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     7081 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     7141 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7501 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7621 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7741 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7981 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8041 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8101 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8161 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8221 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8281 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8341 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8401 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8461 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8521 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8581 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8641 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8701 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8761 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8821 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8881 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8941 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9001 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9061 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     9121 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     9181 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     9241 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     9301 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     9361 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     9421 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     9481 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     9541 gacggccagt gccaagctg
//
