LOCUS       V046060                 9451 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046060
VERSION     V046060
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             2290..3849
                     /label="Lck(NM_001162432)"
                     /note="Lck(NM_001162432)"
                     /gene="Lck"
     CDS             3856..3945
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3994..4205
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4218..4486
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4514..5539
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    5546..6134
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6208..6441
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6513..6634
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6653..6788
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6826..6842)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6850..6866
                     /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(6874..6904)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6919..6940
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7228..7816)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7987..8847)
                     /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(8848..8952)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9426..9442
                     /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 agagccacca tgggggcctc tgagctgacg atctcgggga tcatgggctg tgtctgcagc
     2341 tcaaaccctg aagatgactg gatggagaac attgacgtgt gtgaaaactg ccactatccc
     2401 atagtcccac tggacagcaa gatctcgctg cccatccgga atggctctga agtgcgggac
     2461 ccactggtca cctatgaggg atctctccca ccagcatccc cgctgcaaga caacctggtt
     2521 atcgccctgc acagttatga gccctcccat gatggagact tgggctttga gaagggtgaa
     2581 cagctccgaa tcctggagca gagcggtgag tggtggaagg ctcagtccct gacgactggc
     2641 caagaaggct tcattccctt caacttcgtg gcgaaagcaa acagcctgga gcctgaacct
     2701 tggttcttca agaatctgag ccgtaaggac gccgagcggc agcttttggc gcccgggaac
     2761 acgcatggat ccttcctgat ccgggaaagc gaaagcactg cggggtcctt ttccctgtcg
     2821 gtcagagact tcgaccagaa ccagggagaa gtggtgaaac attacaagat ccgtaaccta
     2881 gacaacggtg gcttctacat ctcccctcgt atcacttttc ccggattgca cgatctagtc
     2941 cgccattaca ccaacgcctc tgatgggctg tgcacaaagt tgagccgtcc ttgccagacc
     3001 cagaagcccc agaaaccatg gtgggaggac gaatgggaag ttcccaggga aacactgaag
     3061 ttggtggagc ggctgggagc tggccagttc ggggaagtgt ggatggggta ctacaacgga
     3121 cacacgaagg tggcggtgaa gagtctgaaa caagggagca tgtcccccga cgccttcctg
     3181 gctgaggcta acctcatgaa gcagctgcag cacccgcggc tagtccggct ttatgcagtg
     3241 gtcacccagg aacccatcta catcatcacg gaatacatgg agaacgggag cctagtagat
     3301 tttctcaaga ctccctcggg catcaagttg aatgtcaaca aacttttgga catggcagcc
     3361 cagattgcag agggcatggc gttcatcgaa gaacagaatt acatccatcg ggacctgcgc
     3421 gccgccaaca tcctggtgtc tgacacgctg agctgcaaga ttgcagactt tggcctggcg
     3481 cgcctcattg aggacaatga gtacacggcc cgggaggggg ccaaatttcc cattaagtgg
     3541 acagcaccag aagccattaa ctatgggacc ttcaccatca agtcagacgt gtggtccttc
     3601 gggatcttgc ttacagagat cgtcacccac ggtcgaatcc cttacccagg aatgaccaac
     3661 cctgaagtca ttcagaacct ggagagaggc taccgcatgg tgagacctga caactgtccg
     3721 gaagagctgt accacctcat gatgctgtgc tggaaggagc gcccagagga ccggcccacg
     3781 tttgactacc ttcggagtgt tctggatgac ttcttcacag ccacagaggg ccagtaccag
     3841 ccccagcctc tcgaggaaca aaaactaata tcagaggaag atttggaaca gaaactaatc
     3901 agtgaagaag atttagagca aaagttaatt tccgaggagg acttatagga tccgcggccg
     3961 caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     4021 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg
     4081 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     4141 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     4201 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     4261 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     4321 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     4381 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     4441 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg
     4501 cgcctacgtc gacatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     4561 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4621 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4681 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     4741 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4801 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4861 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4921 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4981 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     5041 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     5101 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     5161 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     5221 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     5281 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     5341 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     5401 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     5461 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     5521 tccgagggca aaggaatagg tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5581 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5641 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5701 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5761 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5821 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5881 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5941 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6001 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6061 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6121 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6181 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6241 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6301 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6361 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6421 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6481 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6541 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6601 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6661 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6721 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6781 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     6841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9001 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9061 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9241 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9301 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9361 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9421 acgttgtaaa acgacggcca gtgccaagct g
//
