LOCUS       V044490                 9990 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044490
VERSION     V044490
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             3432..4667
                     /label="Dnaja2(NM_019794)"
                     /note="Dnaja2(NM_019794)"
                     /gene="Dnaja2"
     misc_feature    4784..5333
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             5336..5935
                     /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    5959..6547
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6754..7387
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7515..7531)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7539..7555
                     /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(7563..7593)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7608..7629
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7917..8505)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8676..9536)
                     /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(9537..9641)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9689..9823
                     /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 atctgacgcg tatggcgaac gtggccgaca cgaagctgta cgacatcctg ggcgtccctc
     3481 ccggcgctag cgagaacgag ctgaagaagg cataccgaaa gttagccaaa gaataccacc
     3541 ctgataagaa tccaaatgct ggagacaaat ttaaagaaat aagttttgca tatgaagtat
     3601 tgtcaaatcc agagaagcga gagctgtatg acagatatgg agaacaaggc ctacgggaag
     3661 gcagcggcgg aggcggtggc atggatgata tcttctcaca tatttttggt ggaggattgt
     3721 ttggctttat gggcaatcag agtagaagtc gaaatggcag aagaagaggc gaggacatga
     3781 tgcatccact aaaagtatct ttagaagacc tgtacaatgg caagacaacc aaactacaac
     3841 ttagcaagaa tgtgctctgt agtgcatgca gtggccaagg tgggaagtct ggagctgttc
     3901 agaaatgcag cgcttgtcgg ggtcgaggtg tgcgcattat gatcagacag ctggctccag
     3961 gaatggtgca gcagatgcag tccgtgtgct ccgactgtaa tggagaaggg gaggtcatca
     4021 atgaaaaaga ccgctgtaaa aaatgtgaag ggaagaaggt aatcaaagaa gtcaagattc
     4081 tggaagtcca tgtagacaaa ggcatgaaac atggacagag gattacgttc actggggaag
     4141 cagaccaggc tccaggagtg gaacctggag atattgttct tttgctacag gaaaaagaac
     4201 atgaggtgtt ccagagagat gggaatgatt tgcatatgac atataagata ggacttgttg
     4261 aagctttatg tggatttcag ttcacattta aacatcttga tgctcgtcag attgtggtga
     4321 aatacccccc tggcaaagta attgaaccag gatgtgttcg tgttgttcga ggtgaaggaa
     4381 tgccacagta tcgtaatccc tttgaaaagg gtgatctcta cataaagttt gatgtacagt
     4441 ttcctgagaa taactggatc aacccagaca aactttctga attagaagat ctcctgccat
     4501 ctagaccaga agttcctaat gttattggag agacagaaga agtggagctt caggaatttg
     4561 atagcactcg aggctctggc ggtggtcaga gacgtgaagc ctataatgat agctctgatg
     4621 aagaaagtag cagccatcat ggacctggag tgcagtgtgc ccatcagtaa gaattcctcg
     4681 agggcggccg ctctagagtc gacgggccgc ggtaacaatt gttaactaac ttaagctagc
     4741 aacggtttcc ctctagcggg atcaattccg cccccccccc ctaacgttac tggccgaagc
     4801 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     4861 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     4921 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     4981 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     5041 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     5101 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     5161 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     5221 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     5281 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatgac
     5341 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     5401 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     5461 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     5521 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     5581 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     5641 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     5701 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     5761 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5821 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5881 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacccgg
     5941 gcggggcgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6001 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6061 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6121 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6181 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6241 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6301 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     6361 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6421 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6481 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     6541 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta
     6601 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     6661 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     6721 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
     6781 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
     6841 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
     6901 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
     6961 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
     7021 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
     7081 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
     7141 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
     7201 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7261 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7321 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7381 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     7441 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc
     7501 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7561 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7621 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7741 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8521 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8581 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8641 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8701 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8761 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8821 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8881 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8941 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     9001 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     9061 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9121 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     9181 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     9241 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     9301 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9361 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9421 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9481 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9541 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9601 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9661 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa
     9721 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     9781 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
     9841 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg
     9901 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
     9961 ttgttaaata tgtactacaa acttagtagt
//
