LOCUS       V045392                 9361 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045392
VERSION     V045392
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..4191
                     /label="SLC3A2(NM_002394)"
                     /note="SLC3A2(NM_002394)"
                     /gene="SLC3A2"
     CDS             4198..4287
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4331..4542
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4555..4823
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4850..5449
                     /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    5456..6044
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6118..6351
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6423..6544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6563..6698
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6736..6752)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6760..6776
                     /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(6784..6814)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6829..6850
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7138..7726)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7897..8757)
                     /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(8758..8862)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9336..9352
                     /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 catggagcta cagcctcctg aagcctcgat cgccgtcgtg
     2341 tcgattccgc gccagttgcc tggctcacat tcggaggctg gtgtccaggg tctcagcgcg
     2401 ggggacgact cagagttggg gtctcactgt gttgcccaga ctggtctcga actcttggcc
     2461 tcaggtgatc ctcttccctc agcttcccag aatgccgaga tgatagagac ggggtctgac
     2521 tgtgttaccc aggctggtct tcaactcttg gcctcaagtg atcctcctgc cttagcttcc
     2581 aagaatgctg aggttacagg caccatgagc caggacaccg aggtggatat gaaggaggtg
     2641 gagctgaatg agttagagcc cgagaagcag ccgatgaacg cggcgtctgg ggcggccatg
     2701 tccctggcgg gagccgagaa gaatggtctg gtgaagatca aggtggcgga agacgaggcg
     2761 gaggcggcag ccgcggctaa gttcacgggc ctgtccaagg aggagctgct gaaggtggca
     2821 ggcagccccg gctgggtacg cacccgctgg gcactgctgc tgctcttctg gctcggctgg
     2881 ctcggcatgc ttgctggtgc cgtggtcata atcgtgcgag cgccgcgttg tcgcgagcta
     2941 ccggcgcaga agtggtggca cacgggcgcc ctctaccgca tcggcgacct tcaggccttc
     3001 cagggccacg gcgcgggcaa cctggcgggt ctgaaggggc gtctcgatta cctgagctct
     3061 ctgaaggtga agggccttgt gctgggtcca attcacaaga accagaagga tgatgtcgct
     3121 cagactgact tgctgcagat cgaccccaat tttggctcca aggaagattt tgacagtctc
     3181 ttgcaatcgg ctaaaaaaaa gagcatccgt gtcattctgg accttactcc caactaccgg
     3241 ggtgagaact cgtggttctc cactcaggtt gacactgtgg ccaccaaggt gaaggatgct
     3301 ctggagtttt ggctgcaagc tggcgtggat gggttccagg ttcgggacat agagaatctg
     3361 aaggatgcat cctcattctt ggctgagtgg caaaatatca ccaagggctt cagtgaagac
     3421 aggctcttga ttgcggggac taactcctcc gaccttcagc agatcctgag cctactcgaa
     3481 tccaacaaag acttgctgtt gactagctca tacctgtctg attctggttc tactggggag
     3541 catacaaaat ccctagtcac acagtatttg aatgccactg gcaatcgctg gtgcagctgg
     3601 agtttgtctc aggcaaggct cctgacttcc ttcttgccgg ctcaacttct ccgactctac
     3661 cagctgatgc tcttcaccct gccagggacc cctgttttca gctacgggga tgagattggc
     3721 ctggatgcag ctgcccttcc tggacagcct atggaggctc cagtcatgct gtgggatgag
     3781 tccagcttcc ctgacatccc aggggctgta agtgccaaca tgactgtgaa gggccagagt
     3841 gaagaccctg gctccctcct ttccttgttc cggcggctga gtgaccagcg gagtaaggag
     3901 cgctccctac tgcatgggga cttccacgcg ttctccgctg ggcctggact cttctcctat
     3961 atccgccact gggaccagaa tgagcgtttt ctggtagtgc ttaactttgg ggatgtgggc
     4021 ctctcggctg gactgcaggc ctccgacctg cctgccagcg ccagcctgcc agccaaggct
     4081 gacctcctgc tcagcaccca gccaggccgt gaggagggct cccctcttga gctggaacgc
     4141 ctgaaactgg agcctcacga agggctgctg ctccgcttcc cctacgcggc cggatccgaa
     4201 caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga agatttagag
     4261 caaaagttaa tttccgagga ggacttatag cggccgcgaa ggatctgcga tcgctccggt
     4321 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     4381 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     4441 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     4501 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct
     4561 cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt
     4621 cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt
     4681 ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc
     4741 agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt
     4801 tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga tgaccgagta
     4861 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     4921 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     4981 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5041 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5101 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5161 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5221 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5281 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     5341 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5401 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca
     5461 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5521 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5581 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5641 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5701 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5761 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     5821 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     5881 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     5941 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6001 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     6061 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6121 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6181 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6241 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6301 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6361 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6421 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6541 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     6601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6661 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     6721 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6781 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6841 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6901 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6961 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7021 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7081 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7141 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7201 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7261 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7321 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7381 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7441 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7501 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7561 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7621 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7681 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7741 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7801 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7861 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7921 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7981 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8041 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8101 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8161 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8221 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8281 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8341 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8401 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8461 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8521 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8581 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8641 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8701 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8761 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8821 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8881 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8941 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9001 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9061 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9121 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9181 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9241 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9301 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     9361 g
//
