LOCUS       V039249                10270 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039249
VERSION     V039249
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"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2843..4957
                     /label="STT3A(NM_001278503)"
                     /note="STT3A(NM_001278503)"
                     /gene="STT3A"
     CDS             4973..5038
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5058..5557
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5578..6177
                     /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    6191..6779
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6986..7619
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7748..7764)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7772..7788
                     /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(7796..7826)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7841..7862
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8150..8738)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8909..9769)
                     /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(9770..9874)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9922..10056
                     /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 cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattcgcca ccatgactaa gtttggattt ttgcgattgt cctatgagaa
     2881 gcaggacaca cttttgaagc ttctcattct gtcaatggct gctgtattat ccttctccac
     2941 tcgtctgttt gctgtcctga gatttgaaag tgttatccat gagtttgatc cgtactttaa
     3001 ttatcggact accaggttcc tggctgagga ggggttttat aaattccata actggtttga
     3061 tgaccgagcc tggtaccctt tgggacgaat cattggagga acaatttacc caggtttaat
     3121 gatcacctct gctgcaatct accatgtact ccattttttc cacatcacca tcgacattcg
     3181 gaatgtctgt gtgttcctgg cccctctctt ctcctccttc accaccatcg tcacgtacca
     3241 ccttaccaaa gagctcaagg atgcaggggc tgggcttctt gctgctgcca tgattgctgt
     3301 agttcctgga tatatctccc gatctgtggc tggctcctat gataatgaag ggattgccat
     3361 cttttgcatg ctactcacct actacatgtg gatcaaggca gtaaagactg gttccatctg
     3421 ttgggcagct aagtgtgccc ttgcttattt ctacatggtc tcgtcatggg gaggttatgt
     3481 gttcctgatc aacttaattc ctctccacgt cctcgtgctg atgctcacag gccgtttctc
     3541 tcaccggatc tatgtggcct actgtactgt ttactgcctg ggcactatac tttctatgca
     3601 gatctccttt gtgggtttcc agcctgtcct ttcatcagag cacatggcag cctttggggt
     3661 ctttggtctc tgccagatcc atgcctttgt ggattacctg cgcagcaagt tgaatccaca
     3721 acaatttgaa gttcttttcc ggagcgtcat ctctctggta ggctttgtcc ttctcaccgt
     3781 gggagctctc ctcatgctga caggaaaaat atctccctgg acggggcgtt tctactcgct
     3841 gctggatccc tcttatgcta agaacaacat ccccatcatt gcttctgtgt ctgagcatca
     3901 gcccacaacc tggtcctcat actattttga cctgcagctc ctcgtcttca tgtttccagt
     3961 tggcctctat tactgcttta gcaacctgtc tgatgcccgg atttttatca tcatgtatgg
     4021 tgtgaccagc atgtactttt cagctgtaat ggtgcgtcta atgctagtgt tggcacctgt
     4081 tatgtgcatt ctctctggca ttggagtctc ccaggtgctg tccacataca tgaagaatct
     4141 ggacataagt cgtccagaca agaagagcaa gaagcaacag gattccacct accctattaa
     4201 gaatgaagtg gcaagtggga tgatactggt catggctttc tttctcatca cctacacctt
     4261 tcattcaacc tgggtgacca gtgaggccta ctcttctccg tccattgtac tatctgcccg
     4321 tggtggggat ggcagtagga tcatatttga tgacttccga gaagcatatt attggcttcg
     4381 tcataatact ccagaggatg cgaaggtcat gtcctggtgg gattatggct atcagattac
     4441 agctatggca aaccgaacaa ttttagtgga caataacaca tggaataata cccatatttc
     4501 tcgagtaggg caggcaatgg cgtccacaga ggaaaaagcc tatgagatca tgagggagct
     4561 cgatgtcagc tatgtgctgg tcatttttgg aggcctcact gggtattcct ctgatgatat
     4621 caacaagttt ctttggatgg tccggattgg agggagcaca gatacaggca aacatatcaa
     4681 ggagaatgac tattatactc caactgggga gttccgtgtg gaccgtgaag gttctccagt
     4741 gctgctcaac tgcctcatgt acaagatgtg ttactatcgc tttggacagg tttacacaga
     4801 agccaagcgt cctccaggct ttgaccgtgt ccgaaatgct gagattggga ataaagactt
     4861 tgagcttgat gtcctggagg aagcatatac cacagaacat tggctggtca ggatatacaa
     4921 ggtaaaggac ctggataatc gaggcttgtc aaggacaggt ggaggcggat ccgactacaa
     4981 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     5041 gtctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     5101 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     5161 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     5221 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     5281 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     5341 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     5401 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     5461 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     5521 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg
     5581 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     5641 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5701 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5761 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5821 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5881 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5941 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6001 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6061 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6121 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg
     6181 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     6241 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     6301 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6361 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6421 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6481 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     6541 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     6601 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6661 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6721 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6781 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca
     6841 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     6901 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6961 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat
     7021 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca
     7081 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     7141 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg
     7201 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc
     7261 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat
     7321 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg
     7381 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg
     7441 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     7501 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     7561 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     7621 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     7681 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt
     7741 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7801 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7981 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     8041 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     8101 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     8161 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     8221 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     8281 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     8341 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     8401 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     8461 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8521 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8581 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     8641 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8701 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8761 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8821 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8881 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     9001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     9061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     9121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     9181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     9241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     9301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     9361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     9421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     9481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     9541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9901 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc
     9961 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    10021 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
    10081 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
    10141 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
    10201 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa
    10261 acttagtagt
//
