LOCUS       V021831                10146 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021831
VERSION     V021831
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             2301..5000
                     /label="MARS1(NM_004990)"
                     /note="MARS1(NM_004990)"
                     /gene="MARS1"
     CDS             5007..5072
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5116..5327
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5340..5608
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5635..6234
                     /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    6241..6829
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6903..7136
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7208..7329
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7348..7483
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7521..7537)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7545..7561
                     /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(7569..7599)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7614..7635
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7923..8511)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8682..9542)
                     /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(9543..9647)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10121..10137
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgagactgt tcgtgagtga tggcgtcccg ggttgcttgc
     2341 cggtgctggc cgccgccggg agagcccggg gcagagcaga ggtgctcatc agcactgtag
     2401 gcccggaaga ttgtgtggtc ccgttcctga cccggcctaa ggtccctgtc ttgcagctgg
     2461 atagcggcaa ctacctcttc tccactagtg caatctgccg atattttttt ttgttatctg
     2521 gctgggagca agatgacctc actaaccagt ggctggaatg ggaagcgaca gagctgcagc
     2581 cagctttgtc tgctgccctg tactatttag tggtccaagg caagaagggg gaagatgttc
     2641 ttggttcagt gcggagagcc ctgactcaca ttgaccacag cttgagtcgt cagaactgtc
     2701 ctttcctggc tggggagaca gaatctctag ccgacattgt tttgtgggga gccctatacc
     2761 cattactgca agatcccgcc tacctccctg aggagctgag tgccctgcac agctggttcc
     2821 agacactgag tacccaggaa ccatgtcagc gagctgcaga gactgtactg aaacagcaag
     2881 gtgtcctggc tctccggcct tacctccaaa agcagcccca gcccagcccc gctgagggaa
     2941 gggctgtcac caatgagcct gaggaggagg agctggctac cctatctgag gaggagattg
     3001 ctatggctgt tactgcttgg gagaagggcc tagaaagttt gcccccgctg cggccccagc
     3061 agaatccagt gttgcctgtg gctggagaaa ggaatgtgct catcaccagt gccctccctt
     3121 acgtcaacaa tgtcccccac cttgggaaca tcattggttg tgtgctcagt gccgatgtct
     3181 ttgccaggta ctctcgcctc cgccagtgga acaccctcta tctgtgtggg acagatgagt
     3241 atggtacagc aacagagacc aaggctctgg aggagggact aaccccccag gagatctgcg
     3301 acaagtacca catcatccat gctgacatct accgctggtt taacatttcg tttgatattt
     3361 ttggtcgcac caccactcca cagcagacca aaatcaccca ggacattttc cagcagttgc
     3421 tgaaacgagg ttttgtgctg caagatactg tggagcaact gcgatgtgag cactgtgctc
     3481 gcttcctggc tgaccgcttc gtggagggcg tgtgtccctt ctgtggctat gaggaggctc
     3541 ggggtgacca gtgtgacaag tgtggcaagc tcatcaatgc tgtcgagctt aagaagcctc
     3601 agtgtaaagt ctgccgatca tgccctgtgg tgcagtcgag ccagcacctg tttctggacc
     3661 tgcctaagct ggagaagcga ctggaggagt ggttggggag gacattgcct ggcagtgact
     3721 ggacacccaa tgcccagttt atcacccgtt cttggcttcg ggatggcctc aagccacgct
     3781 gcataacccg agacctcaaa tggggaaccc ctgtaccctt agaaggtttt gaagacaagg
     3841 tattctatgt ctggtttgat gccactattg gctatctgtc catcacagcc aactacacag
     3901 accagtggga gagatggtgg aagaacccag agcaagtgga cctgtatcag ttcatggcca
     3961 aagacaatgt tcctttccat agcttagtct ttccttgctc agccctagga gctgaggata
     4021 actatacctt ggtcagccac ctcattgcta cagagtacct gaactatgag gatgggaaat
     4081 tctctaagag ccgcggtgtg ggagtgtttg gggacatggc ccaggacacg gggatccctg
     4141 ctgacatctg gcgcttctat ctgctgtaca ttcggcctga gggccaggac agtgctttct
     4201 cctggacgga cctgctgctg aagaataatt ctgagctgct taacaacctg ggcaacttca
     4261 tcaacagagc tgggatgttt gtgtctaagt tctttggggg ctatgtgcct gagatggtgc
     4321 tcacccctga tgatcagcgc ctgctggccc atgtcaccct ggagctccag cactatcacc
     4381 agctacttga gaaggttcgg atccgggatg ccttgcgcag tatcctcacc atatctcgac
     4441 atggcaacca atatattcag gtgaatgagc cctggaagcg gattaaaggc agtgaggctg
     4501 acaggcaacg ggcaggaaca gtgactggct tggcagtgaa tatagctgcc ttgctctctg
     4561 tcatgcttca gccttacatg cccacggtta gtgccacaat ccaggcccag ctgcagctcc
     4621 cacctccagc ctgcagtatc ctgctgacaa acttcctgtg taccttacca gcaggacacc
     4681 agattggcac agtcagtccc ttgttccaaa aattggaaaa tgaccagatt gaaagtttaa
     4741 ggcagcgctt tggagggggc caggcaaaaa cgtccccgaa gccagcagtt gtagagactg
     4801 ttacaacagc caagccacag cagatacaag cgctgatgga tgaagtgaca aaacaaggaa
     4861 acattgtccg agaactgaaa gcacaaaagg cagacaagaa cgaggttgct gcggaggtgg
     4921 cgaaactctt ggatctaaag aaacagttgg ctgtagctga ggggaaaccc cctgaagccc
     4981 ctaaaggcaa gaagaaaaag ggatccgact acaaagacca tgacggtgat tataaagatc
     5041 atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct
     5101 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     5161 gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     5221 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     5281 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag
     5341 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     5401 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     5461 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     5521 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt
     5581 cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagatgacc
     5641 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc
     5701 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac
     5761 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5821 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5881 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5941 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     6001 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     6061 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     6121 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     6181 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgagtcgac
     6241 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     6301 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     6361 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     6421 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     6481 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     6541 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     6601 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     6661 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     6721 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     6781 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt
     6841 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     6901 actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc
     6961 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     7021 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7081 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     7141 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     7201 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     7261 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     7321 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat
     7381 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     7441 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac
     7501 ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7561 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7621 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7681 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7741 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7801 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7861 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7921 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7981 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     8041 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8101 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8161 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8221 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8281 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8341 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     8401 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8461 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8521 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8581 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8641 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8701 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8761 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8821 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8881 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8941 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     9001 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9061 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9121 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9181 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9241 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9301 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9361 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9421 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9481 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9541 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9601 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9661 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     9721 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     9781 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     9841 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     9901 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     9961 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
    10021 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
    10081 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
    10141 aagctg
//
