LOCUS       V047403                 9451 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047403
VERSION     V047403
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..4305
                     /label="Shcbp1(NM_011369)"
                     /note="Shcbp1(NM_011369)"
                     /gene="Shcbp1"
     CDS             4312..4377
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4421..4632
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4645..4913
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4940..5539
                     /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    5546..6134
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6208..6441
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6513..6634
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6653..6788
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6826..6842)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6850..6866
                     /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(6874..6904)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6919..6940
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7228..7816)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7987..8847)
                     /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(8848..8952)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9426..9442
                     /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 catggctgat gatttgcggg ctggtggagt tctggaacct
     2341 atagctatgg ttccaccgag acctgacttg gcggcggaga aggaaccggc gtcctggaag
     2401 gaaggtttat tcttggatgc agatccatgc agtgatcaag gctatcatgc taatccaggt
     2461 gctactgtaa aaactctcat accagaagga aaaactcctt ttccacgaat tatccaaaca
     2521 aatgaacttc tgttttatga acgattcaga gcctatcaag attacatttt agctgactgt
     2581 aaggcctctg aggtaaagga attcacagtc agcttcttgg aaaaggtcct tgaaccatct
     2641 ggatggtggg cagtctggca cactaatgtg tttgaggtgt tggttgaggt tacaaatgtg
     2701 gactttccat ccctgaaggc ggtcgtaagg cttgcagagc catgcatcta tgaatctaaa
     2761 ttgagcacgt ttaccttggc caatgtgaag gagcttttgg acctgaagga gtttcatctg
     2821 cctctgcagg agttgtgggt ggtatcagat gactcacatg aattccacca gatggcactt
     2881 gcaattgagc acgtcagatt tttctacaaa cacatctgga ggagttggga tgaggaagag
     2941 gaggatgagt atgactattt tgtcagatgt gttgaacctc gactgagatt gtattatgac
     3001 atacttgaag atcgagttcc ctcgggactt attgttgact accacaatct gttgtctcaa
     3061 tgtgaagaga gttacaggaa atttttaaat ctgagaagca gtttgtccaa ttgtaattct
     3121 gattctgagc aggaaaatat ctccatggtg gaagggttaa atttgtattc agaaattgaa
     3181 cagttgaaac aaaagctaaa gctcattgag aatcctttgt taagatatgt ttttggttat
     3241 cagaagaact ctaatatcca aggaaagggt actcgtcaaa atggccagaa ggtcatccat
     3301 gtggtttcct ccaccatgaa gacaggtcta cttcggtctc tattcaagga caggttttgt
     3361 gaggagtctt gcaaagaaga aacagaaatt aagttccata gtgatctgtt gtctggtata
     3421 aatgcctgct atgatggtga cactgtcatt atttgtcctg gccattatgt agttcatggc
     3481 acctgttcca tagctgactc cattgagttg gaaggatatg gcctaccaga tgacattgtc
     3541 atagaaaaga ggggcaaagg agatactttt gtggattgca cgggtatgga tgttaaaatt
     3601 tcaggcataa aatttattca gcatgattct gtggaaggaa tcttaatcat tcaccatggc
     3661 aagaccacac tggaaaactg tgtactacaa tgtgaaacca caggagtcac agtgcgcaca
     3721 tcagcagaac ttttcatgaa aaactcagat gtatatggtg ccaagggtgc tggtatagaa
     3781 atatatcctg gaagtaaatg taccctgact gacaatggaa tccatcactg caaggaagga
     3841 attctcatta aggacttcct tgatgaacat tatgatattc ccaaaatatc gatgataaat
     3901 aacgtcatac acaataatga aggttatggt gttgttttgg tgaagcctac aattttctgt
     3961 gatctacagg aaaatacaca agatgaaatt aatgacaata tggttcagaa aaataaagag
     4021 gcagatgtca ctgaaggatt agatctggaa gaaatgcttc agtgtgtggc tagcaaaatg
     4081 gagccttatg ccacagctga ctttaatgaa caagctaagg gaaactgtga aattataaat
     4141 gaactacttg ctatttccat gcaaaaaggc cggatgaaga aaagactgag tgaacttggg
     4201 attacacaag ctgatgacaa cataatgtca caggagatgt ttattgaaat tatggggaac
     4261 cagtttaagt ggaatggcaa agggagtttt ggcacatttc tttacggatc cgactacaaa
     4321 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     4381 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc
     4441 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     4501 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     4561 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     4621 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     4681 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     4741 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     4801 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     4861 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     4921 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     4981 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5041 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5101 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5161 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5221 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5281 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5341 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5401 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5461 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5521 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5581 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5641 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5701 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5761 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5821 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5881 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5941 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6001 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6061 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6121 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6181 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6241 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6301 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6361 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6421 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6481 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6661 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6721 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6781 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     6841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9001 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9061 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9241 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9301 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9361 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9421 acgttgtaaa acgacggcca gtgccaagct g
//
