LOCUS       V035139                 9241 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035139
VERSION     V035139
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..4095
                     /label="NR4A1(NM_002135)"
                     /note="NR4A1(NM_002135)"
                     /gene="NR4A1"
     CDS             4102..4167
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4211..4422
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4435..4703
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4730..5329
                     /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    5336..5924
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5998..6231
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6303..6424
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6443..6578
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6616..6632)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6640..6656
                     /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(6664..6694)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6709..6730
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7018..7606)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7777..8637)
                     /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(8638..8742)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9216..9232
                     /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 catgccctgt atccaagccc aatatgggac accagcaccg
     2341 agtccgggac cccgtgacca cctggcaagc gaccccctga cccctgagtt catcaagccc
     2401 accatggacc tggccagccc cgaggcagcc cccgctgccc ccactgccct gcccagcttc
     2461 agcaccttca tggacggcta cacaggagag tttgacacct tcctctacca gctgccagga
     2521 acagtccagc catgctcctc agcctcctcc tcggcctcct ccacatcctc gtcctcagcc
     2581 acctcccctg cctctgcctc cttcaagttc gaggacttcc aggtgtacgg ctgctacccc
     2641 ggccccctga gcggcccagt ggatgaggcc ctgtcctcca gtggctctga ctactatggc
     2701 agcccctgct cggccccgtc gccctccacg cccagcttcc agccgcccca gctctctccc
     2761 tgggatggct ccttcggcca cttctcgccc agccagactt acgaaggcct gcgggcatgg
     2821 acagagcagc tgcccaaagc ctctgggccc ccacagcctc cagccttctt ttccttcagt
     2881 cctcccaccg gccccagccc cagcctggcc cagagccccc tgaagttgtt cccctcacag
     2941 gccacccacc agctggggga gggagagagc tattccatgc ctacggcctt cccaggtttg
     3001 gcacccactt ctccacacct tgagggctcg gggatactgg atacacccgt gacctcaacc
     3061 aaggcccgga gcggggcccc aggtggaagt gaaggccgct gtgctgtgtg tggggacaac
     3121 gcttcatgcc agcattatgg tgtccgcaca tgtgagggct gcaagggctt cttcaagcgc
     3181 acagtgcaga aaaacgccaa gtacatctgc ctggctaaca aggactgccc tgtggacaag
     3241 aggcggcgaa accgctgcca gttctgccgc ttccagaagt gcctggcggt gggcatggtg
     3301 aaggaagttg tccgaacaga cagcctgaag gggcggcggg gccggctacc ttcaaaaccc
     3361 aagcagcccc cagatgcctc ccctgccaat ctcctcactt ccctggtccg tgcacacctg
     3421 gactcagggc ccagcactgc caaactggac tactccaagt tccaggagct ggtgctgccc
     3481 cactttggga aggaagatgc tggggatgta cagcagttct acgacctgct ctccggttct
     3541 ctggaggtca tccgcaagtg ggcggagaag atccctggct ttgctgagct gtcaccggct
     3601 gaccaggacc tgttgctgga gtcggccttc ctggagctct tcatcctccg cctggcgtac
     3661 aggtctaagc caggcgaggg caagctcatc ttctgctcag gcctggtgct acaccggctg
     3721 cagtgtgccc gtggcttcgg ggactggatt gacagtatcc tggccttctc aaggtccctg
     3781 cacagcttgc ttgtcgatgt ccctgccttc gcctgcctct ctgcccttgt cctcatcacc
     3841 gaccggcatg ggctgcagga gccgcggcgg gtggaggagc tgcagaaccg catcgccagc
     3901 tgcctgaagg agcacgtggc agctgtggcg ggcgagcccc agccagccag ctgcctgtca
     3961 cgtctgttgg gcaaactgcc cgagctgcgg accctgtgca cccagggcct gcagcgcatc
     4021 ttctacctca agctggagga cttggtgccc cctccaccca tcattgacaa gatcttcatg
     4081 gacacgctgc ccttcggatc cgactacaaa gaccatgacg gtgattataa agatcatgac
     4141 atcgactaca aggatgacga tgacaagtag cggccgcgaa ggatctgcga tcgctccggt
     4201 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     4261 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     4321 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     4381 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     4441 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     4501 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     4561 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     4621 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     4681 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga tgaccgagta
     4741 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     4801 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     4861 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4921 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4981 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5041 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5101 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5161 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     5221 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5281 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca
     5341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5461 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5521 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5581 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5641 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5701 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     5761 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5821 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     5881 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     5941 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6001 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6061 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6121 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6181 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6241 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6301 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6361 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6421 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     6481 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6541 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     6601 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6661 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6721 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6781 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7441 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7741 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7801 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7861 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7921 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7981 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8041 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8101 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8161 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8221 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8281 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8341 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8401 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8461 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8521 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8581 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8701 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8761 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8821 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     8881 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     8941 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9001 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9061 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9121 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9181 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     9241 g
//
