LOCUS       V041495                 9660 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041495
VERSION     V041495
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     promoter        2221..2432
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     intron          2433..3371
                     /note="intron upstream of the start codon of human EF-1α"
                     /label="EF-1α intron A"
     regulatory      3387..3396
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3396..3425
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3426..4337
                     /label="Stub1(NM_019719)"
                     /note="Stub1(NM_019719)"
                     /gene="Stub1"
     misc_feature    4454..5003
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5006..5605
                     /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    5629..6217
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6424..7057
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7185..7201)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7209..7225
                     /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(7233..7263)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7278..7299
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7587..8175)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8346..9206)
                     /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(9207..9311)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9359..9493
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgaggt gcccgtcagt
     2221 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2281 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2341 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2401 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2461 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     2521 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     2581 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     2641 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     2701 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     2761 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     2821 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     2881 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     2941 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     3001 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     3061 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     3121 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     3181 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     3241 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     3301 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     3361 ttccatttca ggtgtcgtga ggatccgcca ccatggaaca aaaactcatc tcagaagagg
     3421 atctgatgaa gggcaaggag gaaaaggagg gcggcgcgcg gctgggcact ggtggcggcg
     3481 gcagccctga taagagcccg agtgcgcaag agctcaagga gcagggaaac cggctcttcg
     3541 tgggccgcaa gtacccggag gcggcggcct gctacggccg cgccatcact cggaacccac
     3601 ttgtggcagt gtactacact aaccgggccc tgtgctatct gaagatgcag cagcctgaac
     3661 aggcacttgc tgactgccgg cgagccctgg agctggacgg gcagtctgtg aaggcgcact
     3721 tcttcctggg gcagtgccag ctggagatgg agagttatga tgaggccatt gccaatctgc
     3781 agcgagccta tagtttggcc aaggagcagc gactcaactt tggggatgat attcctagtg
     3841 cccttcgcat tgctaagaag aagcgctgga acagtatcga ggaacggcgc atccaccagg
     3901 agagtgagct gcattcatat ctcaccaggc tcattgctgc tgagcgagag agggaactgg
     3961 aggagtgtca gcggaaccac gagggtcatg aagatgatgg ccacatccgg gcccagcagg
     4021 cctgcattga ggccaagcac gataaataca tggcagatat ggatgagctc ttctctcagg
     4081 tggacgagaa aagaaagaag cgagatatcc ctgactactt gtgtggcaag attagctttg
     4141 agctgatgcg ggaaccctgc attacaccca gtggtatcac ctatgaccgc aaggacattg
     4201 aggagcacct gcagcgtgtg ggccactttg accctgtgac ccggagccct ctgacccagg
     4261 aacagctcat ccccaacttg gccatgaagg aagtcattga cgctttcatc tctgagaacg
     4321 gctgggtaga ggactattga gaattcctcg agggcggccg ctctagagtc gacgggccgc
     4381 ggtaacaatt gttaactaac ttaagctagc aacggtttcc ctctagcggg atcaattccg
     4441 cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     4501 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     4561 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     4621 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     4681 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     4741 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     4801 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     4861 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     4921 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     4981 ttcctttgaa aaacacgata ataccatgac cgagtacaag cccacggtgc gcctcgccac
     5041 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5101 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5161 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5221 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5281 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5341 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5401 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5461 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5521 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5581 catgacccgc aagcccggtg cctgacccgg gcggggcgcg tctggaacaa tcaacctctg
     5641 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5701 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5761 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5821 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5881 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5941 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6001 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     6061 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6121 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6181 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     6241 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     6301 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     6361 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg
     6421 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     6481 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     6541 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     6601 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     6661 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     6721 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     6781 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     6841 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     6901 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6961 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7021 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     7081 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc
     7141 tagcgtttac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7201 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7261 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7321 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7381 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8461 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8641 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8941 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9181 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9241 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9301 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa
     9361 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     9421 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9481 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     9541 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     9601 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
