LOCUS       V035217                 9571 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035217
VERSION     V035217
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..4425
                     /label="FBXO43(NM_001029860)"
                     /note="FBXO43(NM_001029860)"
                     /gene="FBXO43"
     CDS             4432..4497
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4541..4752
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4765..5033
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5060..5659
                     /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    5666..6254
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6328..6561
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6633..6754
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6773..6908
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6946..6962)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6970..6986
                     /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(6994..7024)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7039..7060
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7348..7936)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8107..8967)
                     /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(8968..9072)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9546..9562
                     /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 catgagtttt aaagacaaag atgagagaat ttcttgtttg
     2341 gaagcctacg taactttgac atctaagagc tcaagattta ctgatgaaac agagattttg
     2401 aagatgtcgc aaaggcactc aggtcaagct ggcactgaag caggaaatgg ggcggactct
     2461 cctccaattg tcaactccaa gtactccacc ttcagagatt tttgttccac atcttcattt
     2521 caagatagtg gctacaatga gttaaaatct tgtagctttg ataatataga taaagaatat
     2581 cttggaaaga aagaaaaagg cccaacatta ctctatgagc accctgaaac ttcaggcctg
     2641 ggcttaacac atcctttaga atctcccact caaaaaaaga aatgtatctt gcctagaaag
     2701 gaaaaggata aaaccccaga actttgtgaa acacctaaaa tcagtgggaa aaaatgttta
     2761 cctcgcagaa ggttgaatgt atctttcgct cttctaaaag gggactttga atcacaaaat
     2821 agttctttag aaagtagtat aagccaagtt atcaacttag aaaaaaatat tccaagcagt
     2881 gcttcaggtt tttccagggc aaataatttt agccctttag ttactagcac tttaaaaaca
     2941 gaagaagtga cttcatgcag tcaaaaattg aggcttaatt tttctcagca aaagacttcc
     3001 acaattgatg attccaaaga tgattgtagc ctatttgaag ttgaatgtat atctccaatt
     3061 cagggcaata attttaaaga ctctatcaca catgacttta gtgatagcag tttatgcatt
     3121 aatgatgaga atgcatgtcc agagctcctg ggctcctctg ttagtggaac aacttgtgga
     3181 acagatgagg acatatttgt gactccgata agtaatcttg tggcaaacat tagatttaac
     3241 gcaagtcaaa tactttctcc ttcacctgaa gtgagaggca gtatttcaac gcctgaagac
     3301 agtggtttta actcacttag cttggagaaa tcagaagatt ccctgtctga ccaggagggt
     3361 tcttttcaag aactactgca gaaacataag gggactccca aagttgggga caccataaga
     3421 aagacaagac atcttggaag gtcgagaaga ctgtccaccc ttcgggaaca aagctcgcag
     3481 tcagagacag aagaggaaaa gcagattgtc caccctgact ctgaaaaaag agcagcagct
     3541 gcttctgcca tctcagaggg tcagctgagt agtgatgaga gtggggattt gacctttagc
     3601 ttaaagaatt tatcaaagac cccagccttg caattggtac atgagctgtt catgaaaagc
     3661 aagaggaaaa gattacagga aaatagtgga catgaattct tagagcaagg ggatggggag
     3721 aaaatagctg tactgcagtg tatacttgca ggactgatcg gcaagaaaat gggtatagaa
     3781 aaactggaca tcttaacaga attaaaatat agaaatttaa agcatattct tgctatggtt
     3841 ttagagtcct tgaccgcaga gagcctatgc agtgtttgga aagtaagcag aaattggcgt
     3901 gaaattgttg ttcaagataa aaatgcaaat cggaggagga aattttatat cacacaactg
     3961 aaaacagatt ctgagggggc tgtattaaat gtcgaggatg ctgccactcg gctccagctt
     4021 ttaaatcgct cagctttaag atctgtgcag gcacaggcta ggatacctgg ttctcagaga
     4081 gagcaagggt caacattatc tccctgggga gaagttttga cacctctagc aagctcttct
     4141 gttactcact taagtagtaa acaggaagaa tatgttaagg ttgccaaaac actttttact
     4201 gatgaagcat taaaaccttg cccaaggtgc cagtcccctg ctaagtacca gccatataag
     4261 aaaaggggac tgtgtagccg aacagcctgt ggttttgact tttgtgtgtt atgtctgtgt
     4321 gcttatcatg ggtctgaaga atgtagtaga ggagcagcaa agccaagaaa tagaaaagat
     4381 gctctcccag gaagtgccca gagtaagcgg aatttaaaac gcctcggatc cgactacaaa
     4441 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     4501 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc
     4561 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     4621 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     4681 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     4741 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     4801 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     4861 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     4921 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     4981 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     5041 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5101 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5161 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5221 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5281 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5341 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5401 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5461 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5521 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5581 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5641 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5701 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5761 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5821 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5881 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5941 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     6001 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     6061 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6121 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6181 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6241 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6301 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6361 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6421 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6481 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6541 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6601 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6661 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6721 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6781 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6841 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6901 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     6961 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7021 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7141 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7201 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7261 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7321 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7381 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7441 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7501 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7561 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7621 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7681 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7741 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7801 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7861 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7921 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7981 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8041 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8101 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8221 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8401 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9061 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9121 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9181 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9241 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9301 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9361 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9421 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9481 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9541 acgttgtaaa acgacggcca gtgccaagct g
//
