LOCUS       V029125                 8841 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029125
VERSION     V029125
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"
     regulatory      2288..2297
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2297..2362
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2369..3766
                     /label="GCK(NM_033507)"
                     /note="GCK(NM_033507)"
                     /gene="GCK"
     promoter        3811..4022
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4035..4303
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4330..4929
                     /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    4936..5524
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5598..5831
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5903..6024
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6043..6178
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6216..6232)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6240..6256
                     /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(6264..6294)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6309..6330
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6618..7206)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7377..8237)
                     /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(8238..8342)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8816..8832
                     /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2341 ctacaaggat gacgatgaca aggaattcat ggcgatggat gtcacaagga gccaggccca
     2401 gacagccttg actctggtag agcagatcct ggcagagttc cagctgcagg aggaggacct
     2461 gaagaaggtg atgagacgga tgcagaagga gatggaccgc ggcctgaggc tggagaccca
     2521 tgaagaggcc agtgtgaaga tgctgcccac ctacgtgcgc tccaccccag aaggctcaga
     2581 agtcggggac ttcctctccc tggacctggg tggcactaac ttcagggtga tgctggtgaa
     2641 ggtgggagaa ggtgaggagg ggcagtggag cgtgaagacc aaacaccaga tgtactccat
     2701 ccccgaggac gccatgaccg gcactgctga gatgctcttc gactacatct ctgagtgcat
     2761 ctccgacttc ctggacaagc atcagatgaa acacaagaag ctgcccctgg gcttcacctt
     2821 ctcctttcct gtgaggcacg aagacatcga taagggcatc cttctcaact ggaccaaggg
     2881 cttcaaggcc tcaggagcag aagggaacaa tgtcgtgggg cttctgcgag acgctatcaa
     2941 acggagaggg gactttgaaa tggatgtggt ggcaatggtg aatgacacgg tggccacgat
     3001 gatctcctgc tactacgaag accatcagtg cgaggtcggc atgatcgtgg gcacgggctg
     3061 caatgcctgc tacatggagg agatgcagaa tgtggagctg gtggaggggg acgagggccg
     3121 catgtgcgtc aataccgagt ggggcgcctt cggggactcc ggcgagctgg acgagttcct
     3181 gctggagtat gaccgcctgg tggacgagag ctctgcaaac cccggtcagc agctgtatga
     3241 gaagctcata ggtggcaagt acatgggcga gctggtgcgg cttgtgctgc tcaggctcgt
     3301 ggacgaaaac ctgctcttcc acggggaggc ctccgagcag ctgcgcacac gcggagcctt
     3361 cgagacgcgc ttcgtgtcgc aggtggagag cgacacgggc gaccgcaagc agatctacaa
     3421 catcctgagc acgctggggc tgcgaccctc gaccaccgac tgcgacatcg tgcgccgcgc
     3481 ctgcgagagc gtgtctacgc gcgctgcgca catgtgctcg gcggggctgg cgggcgtcat
     3541 caaccgcatg cgcgagagcc gcagcgagga cgtaatgcgc atcactgtgg gcgtggatgg
     3601 ctccgtgtac aagctgcacc ccagcttcaa ggagcggttc catgccagcg tgcgcaggct
     3661 gacgcccagc tgcgagatca ccttcatcga gtcggaggag ggcagtggcc ggggcgcggc
     3721 cctggtctcg gcggtggcct gtaagaaggc ctgtatgctg ggccagtgag cggccgcgaa
     3781 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     3841 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
     3901 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     3961 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     4021 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
     4081 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
     4141 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
     4201 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
     4261 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc
     4321 ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4381 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4441 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4501 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4561 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     4621 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     4681 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     4741 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     4801 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     4861 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4921 ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4981 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     5041 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5101 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5161 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5221 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5281 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5341 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5401 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5461 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5521 ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     5581 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt
     5641 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     5701 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5761 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     5821 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     5881 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     5941 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6001 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     6061 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6121 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6181 gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6241 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6301 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6361 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6421 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6481 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6541 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6601 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6661 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6721 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6781 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6841 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6961 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7021 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7081 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7141 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7201 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7261 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7321 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7381 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7441 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7501 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7561 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7621 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7681 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7741 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7801 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7861 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7921 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7981 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8041 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8101 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8161 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8221 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8281 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8341 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     8401 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     8461 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     8521 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     8581 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     8641 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     8701 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     8761 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     8821 acgacggcca gtgccaagct g
//
