LOCUS       V039194                 9706 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039194
VERSION     V039194
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..4536
                     /label="CHUK(NM_001278)"
                     /note="CHUK(NM_001278)"
                     /gene="CHUK"
     CDS             4543..4632
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4676..4887
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4900..5168
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5195..5794
                     /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    5801..6389
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6463..6696
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6768..6889
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6908..7043
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7081..7097)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7105..7121
                     /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(7129..7159)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7174..7195
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7483..8071)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8242..9102)
                     /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(9103..9207)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9681..9697
                     /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 catggagcgg cccccggggc tgcggccggg cgcgggcggg
     2341 ccctgggaga tgcgggagcg gctgggcacc ggcggcttcg ggaacgtctg tctgtaccag
     2401 catcgggaac ttgatctcaa aatagcaatt aagtcttgtc gcctagagct aagtaccaaa
     2461 aacagagaac gatggtgcca tgaaatccag attatgaaga agttgaacca tgccaatgtt
     2521 gtaaaggcct gtgatgttcc tgaagaattg aatattttga ttcatgatgt gcctcttcta
     2581 gcaatggaat actgttctgg aggagatctc cgaaagctgc tcaacaaacc agaaaattgt
     2641 tgtggactta aagaaagcca gatactttct ttactaagtg atatagggtc tgggattcga
     2701 tatttgcatg aaaacaaaat tatacatcga gatctaaaac ctgaaaacat agttcttcag
     2761 gatgttggtg gaaagataat acataaaata attgatctgg gatatgccaa agatgttgat
     2821 caaggaagtc tgtgtacatc ttttgtggga acactgcagt atctggcccc agagctcttt
     2881 gagaataagc cttacacagc cactgttgat tattggagct ttgggaccat ggtatttgaa
     2941 tgtattgctg gatataggcc ttttttgcat catctgcagc catttacctg gcatgagaag
     3001 attaagaaga aggatccaaa gtgtatattt gcatgtgaag agatgtcagg agaagttcgg
     3061 tttagtagcc atttacctca accaaatagc ctttgtagtt tagtagtaga acccatggaa
     3121 aactggctac agttgatgtt gaattgggac cctcagcaga gaggaggacc tgttgacctt
     3181 actttgaagc agccaagatg ttttgtatta atggatcaca ttttgaattt gaagatagta
     3241 cacatcctaa atatgacttc tgcaaagata atttcttttc tgttaccacc tgatgaaagt
     3301 cttcattcac tacagtctcg tattgagcgt gaaactggaa taaatactgg ttctcaagaa
     3361 cttctttcag agacaggaat ttctctggat cctcggaaac cagcctctca atgtgttcta
     3421 gatggagtta gaggctgtga tagctatatg gtttatttgt ttgataaaag taaaactgta
     3481 tatgaagggc catttgcttc cagaagttta tctgattgtg taaattatat tgtacaggac
     3541 agcaaaatac agcttccaat tatacagctg cgtaaagtgt gggctgaagc agtgcactat
     3601 gtgtctggac taaaagaaga ctatagcagg ctctttcagg gacaaagggc agcaatgtta
     3661 agtcttctta gatataatgc taacttaaca aaaatgaaga acactttgat ctcagcatca
     3721 caacaactga aagctaaatt ggagtttttt cacaaaagca ttcagcttga cttggagaga
     3781 tacagcgagc agatgacgta tgggatatct tcagaaaaaa tgctaaaagc atggaaagaa
     3841 atggaagaaa aggccatcca ctatgctgag gttggtgtca ttggatacct ggaggatcag
     3901 attatgtctt tgcatgctga aatcatggag ctacagaaga gcccctatgg aagacgtcag
     3961 ggagacttga tggaatctct ggaacagcgt gccattgatc tatataagca gttaaaacac
     4021 agaccttcag atcactccta cagtgacagc acagagatgg tgaaaatcat tgtgcacact
     4081 gtgcagagtc aggaccgtgt gctcaaggag ctgtttggtc atttgagcaa gttgttgggc
     4141 tgtaagcaga agattattga tctactccct aaggtggaag tggccctcag taatatcaaa
     4201 gaagctgaca atactgtcat gttcatgcag ggaaaaaggc agaaagaaat atggcatctc
     4261 cttaaaattg cctgtacaca gagttctgcc cggtcccttg taggatccag tctagaaggt
     4321 gcagtaaccc ctcagacatc agcatggctg cccccgactt cagcagaaca tgatcattct
     4381 ctgtcatgtg tggtaactcc tcaagatggg gagacttcag cacaaatgat agaagaaaat
     4441 ttgaactgcc ttggccattt aagcactatt attcatgagg caaatgagga acagggcaat
     4501 agtatgatga atcttgattg gagttggtta acagaaggat ccgaacaaaa actaatatca
     4561 gaggaagatt tggaacagaa actaatcagt gaagaagatt tagagcaaaa gttaatttcc
     4621 gaggaggact tatagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     4681 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     4741 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     4801 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     4861 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     4921 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     4981 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     5041 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     5101 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     5161 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg
     5221 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     5281 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     5341 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5401 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5461 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5521 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5581 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5641 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5701 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5761 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa
     5821 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5881 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5941 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6001 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     6061 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6121 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6181 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     6241 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6301 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6361 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     6421 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6481 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6541 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6601 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6661 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6721 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6781 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6841 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6901 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6961 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     7021 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     7081 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7141 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7201 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7261 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7921 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     9241 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     9301 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     9361 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     9421 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     9481 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     9541 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     9601 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     9661 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
