LOCUS       V042389                 9751 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042389
VERSION     V042389
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..4581
                     /label="TFRC(NM_003234)"
                     /note="TFRC(NM_003234)"
                     /gene="TFRC"
     CDS             4588..4677
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4721..4932
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4945..5213
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5240..5839
                     /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    5846..6434
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6508..6741
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6813..6934
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6953..7088
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7126..7142)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7150..7166
                     /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(7174..7204)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7219..7240
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7528..8116)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8287..9147)
                     /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(9148..9252)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9726..9742
                     /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 catgatggat caagctagat cagcattctc taacttgttt
     2341 ggtggagaac cattgtcata tacccggttc agcctggctc ggcaagtaga tggcgataac
     2401 agtcatgtgg agatgaaact tgctgtagat gaagaagaaa atgctgacaa taacacaaag
     2461 gccaatgtca caaaaccaaa aaggtgtagt ggaagtatct gctatgggac tattgctgtg
     2521 atcgtctttt tcttgattgg atttatgatt ggctacttgg gctattgtaa aggggtagaa
     2581 ccaaaaactg agtgtgagag actggcagga accgagtctc cagtgaggga ggagccagga
     2641 gaggacttcc ctgcagcacg tcgcttatat tgggatgacc tgaagagaaa gttgtcggag
     2701 aaactggaca gcacagactt caccggcacc atcaagctgc tgaatgaaaa ttcatatgtc
     2761 cctcgtgagg ctggatctca aaaagatgaa aatcttgcgt tgtatgttga aaatcaattt
     2821 cgtgaattta aactcagcaa agtctggcgt gatcaacatt ttgttaagat tcaggtcaaa
     2881 gacagcgctc aaaactcggt gatcatagtt gataagaacg gtagacttgt ttacctggtg
     2941 gagaatcctg ggggttatgt ggcgtatagt aaggctgcaa cagttactgg taaactggtc
     3001 catgctaatt ttggtactaa aaaagatttt gaggatttat acactcctgt gaatggatct
     3061 atagtgattg tcagagcagg gaaaatcacc tttgcagaaa aggttgcaaa tgctgaaagc
     3121 ttaaatgcaa ttggtgtgtt gatatacatg gaccagacta aatttcccat tgttaacgca
     3181 gaactttcat tctttggaca tgctcatctg gggacaggtg acccttacac acctggattc
     3241 ccttccttca atcacactca gtttccacca tctcggtcat caggattgcc taatatacct
     3301 gtccagacaa tctccagagc tgctgcagaa aagctgtttg ggaatatgga aggagactgt
     3361 ccctctgact ggaaaacaga ctctacatgt aggatggtaa cctcagaaag caagaatgtg
     3421 aagctcactg tgagcaatgt gctgaaagag ataaaaattc ttaacatctt tggagttatt
     3481 aaaggctttg tagaaccaga tcactatgtt gtagttgggg cccagagaga tgcatggggc
     3541 cctggagctg caaaatccgg tgtaggcaca gctctcctat tgaaacttgc ccagatgttc
     3601 tcagatatgg tcttaaaaga tgggtttcag cccagcagaa gcattatctt tgccagttgg
     3661 agtgctggag actttggatc ggttggtgcc actgaatggc tagagggata cctttcgtcc
     3721 ctgcatttaa aggctttcac ttatattaat ctggataaag cggttcttgg taccagcaac
     3781 ttcaaggttt ctgccagccc actgttgtat acgcttattg agaaaacaat gcaaaatgtg
     3841 aagcatccgg ttactgggca atttctatat caggacagca actgggccag caaagttgag
     3901 aaactcactt tagacaatgc tgctttccct ttccttgcat attctggaat cccagcagtt
     3961 tctttctgtt tttgcgagga cacagattat ccttatttgg gtaccaccat ggacacctat
     4021 aaggaactga ttgagaggat tcctgagttg aacaaagtgg cacgagcagc tgcagaggtc
     4081 gctggtcagt tcgtgattaa actaacccat gatgttgaat tgaacctgga ctatgagagg
     4141 tacaacagcc aactgctttc atttgtgagg gatctgaacc aatacagagc agacataaag
     4201 gaaatgggcc tgagtttaca gtggctgtat tctgctcgtg gagacttctt ccgtgctact
     4261 tccagactaa caacagattt cgggaatgct gagaaaacag acagatttgt catgaagaaa
     4321 ctcaatgatc gtgtcatgag agtggagtat cacttcctct ctccctacgt atctccaaaa
     4381 gagtctcctt tccgacatgt cttctggggc tccggctctc acacgctgcc agctttactg
     4441 gagaacttga aactgcgtaa acaaaataac ggtgctttta atgaaacgct gttcagaaac
     4501 cagttggctc tagctacttg gactattcag ggagctgcaa atgccctctc tggtgacgtt
     4561 tgggacattg acaatgagtt tggatccgaa caaaaactaa tatcagagga agatttggaa
     4621 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatag
     4681 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc
     4741 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     4801 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     4861 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     4921 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     4981 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     5041 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     5101 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     5161 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     5221 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5281 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5341 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5401 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5461 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5521 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5581 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5641 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5701 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5761 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5821 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5881 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5941 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     6001 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     6061 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     6121 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     6181 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     6241 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6301 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6361 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6421 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6481 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6541 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6601 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6661 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6721 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6781 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6841 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6901 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6961 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     7021 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7081 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     7141 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7201 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7261 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7321 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9241 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9301 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9361 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9421 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9481 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9541 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9601 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9661 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9721 acgttgtaaa acgacggcca gtgccaagct g
//
