LOCUS       V019603                 9660 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019603
VERSION     V019603
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             2807..3514
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             3548..4453
                     /label="STX17(NM_017919)"
                     /note="STX17(NM_017919)"
                     /gene="STX17"
     promoter        4495..4994
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5015..5614
                     /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    5628..6216
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6423..7056
                     /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 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 tctaccggtc gccaccatgg tgagcaaggg
     2821 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2881 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2941 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3001 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3061 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3121 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3181 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     3241 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3301 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     3361 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     3421 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     3481 ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc gaggaggagg
     3541 gggaagcatg tctgaagatg aagaaaaagt gaaattacgc cgtcttgaac cagctatcca
     3601 gaaattcatt aagatagtaa tcccaacaga cctggaaagg ttaagaaagc accagataaa
     3661 tattgagaag tatcaaaggt gcagaatctg ggacaagttg catgaagagc atatcaatgc
     3721 aggacgtaca gttcagcaac tccgatccaa tatccgagaa attgagaaac tttgtttgaa
     3781 agtccgaaag gatgacctag tacttctgaa gagaatgata gatcctgtta aagaagaagc
     3841 atcagcagca acagcagaat ttctccaact ccatttggaa tctgtagaag aacttaagaa
     3901 gcaatttaat gatgaagaaa ctttgctaca gcctcctttg accagatcca tgactgttgg
     3961 tggagcattt catactactg aagctgaagc tagttctcag agtttgactc agatatatgc
     4021 cttacctgaa attcctcaag atcaaaatgc tgcagaatcg tgggaaacct tagaagcgga
     4081 cttaattgaa cttagccaac tggtcactga cttctctctc ctagtgaatt ctcagcagga
     4141 gaagattgac agcattgcag accatgtcaa cagtgctgct gtgaatgttg aagagggaac
     4201 caaaaactta gggaaggctg caaaatacaa gctggcagct ctgcctgtgg caggtgcact
     4261 catcggggga atggtagggg gtcctattgg cctccttgca ggcttcaaag tggcaggaat
     4321 tgcagctgca cttggtggtg gggtgttggg cttcacaggt ggaaaattga tacaaagaaa
     4381 gaaacagaaa atgatggaga agctcacttc cagctgtcca gatcttccca gccaaactga
     4441 caagaaatgc agttagggat ccaccggatc tagataactg atcataattc taccgggtag
     4501 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     4561 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     4621 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     4681 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     4741 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     4801 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     4861 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     4921 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     4981 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     5041 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     5101 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     5161 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5221 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5281 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5341 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5401 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5461 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5521 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5581 cacctggtgc atgacccgca agcccggtgc ctgaccgcgt ctggaacaat caacctctgg
     5641 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5701 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5761 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5821 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5881 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5941 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6001 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     6061 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6121 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6181 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
     6241 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     6301 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     6361 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     6421 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     6481 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     6541 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     6601 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     6661 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     6721 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     6781 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     6841 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     6901 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6961 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     7021 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     7081 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     7141 agcgttttac 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
//
