LOCUS       V028107                 9485 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028107
VERSION     V028107
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             2826..4103
                     /label="PSG8(NM_182707)"
                     /note="PSG8(NM_182707)"
                     /gene="PSG8"
     CDS             4128..4211
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     promoter        4273..4772
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4793..5392
                     /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    5406..5994
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6201..6834
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6963..6979)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6987..7003
                     /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(7011..7041)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7056..7077
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7365..7953)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8124..8984)
                     /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(8985..9089)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9137..9271
                     /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 ccaccatggg gctcctctca gcccctccct gcacacagcg catcacctgg aaggggctcc
     2881 tgctcacagc atcactttta aacttctgga acccacccac gactgcccaa gtcacgattg
     2941 aagcccagcc aaccaaagtt tctgagggga aggatgttct tctacttgtc cacaatttgc
     3001 cccagaatct tactggctac atctggtaca aagggcaaat cagggacctc taccattaca
     3061 ttacatcata tgtagtagac ggtcaaataa ttatatatgg gcctgcatac agtggacgag
     3121 aaacaatata ttccaatgca tccctgctga tccagaatgt cacccaggaa gacgcaggat
     3181 cctacacctt acacatcata atgggaggtg atgagaatag aggagtaact ggacatttca
     3241 ccttcacctt atatctggag actcccaagc cctccatctc cagcagcaaa ttaaacccca
     3301 gggaggccat ggaggctgtg agcttaacct gtgatcctga gactccggac gcaagctacc
     3361 tgtggtggat gaatggtcag agcctcccta tgtctcacag gttgcagttg tctgaaacca
     3421 acaggaccct ctttctattg ggtgtcacaa agtacactgc aggaccctat gaatgtgaaa
     3481 tacggaaccc agtgagtgcc agccgcagtg acccattcac cctgaatctc ctcccgaagc
     3541 tgcccaagcc ctacatcacc atcaacaact taaaacccag ggagaataag gatgtcttaa
     3601 acttcacctg tgaacctaag agtgagaact acacctacat ttggtggcta aatggtcaga
     3661 gcctcccggt cagtcccagg gtaaagcgac ccattgaaaa caggatcctc attctaccca
     3721 gtgtcacgag aaatgaaaca ggaccctatc aatgtgaaat aagggaccaa tatggtggca
     3781 tccgcagtta cccagtcacc ctgaatgtcc tctatggtcc agacctcccc agaatttacc
     3841 cttcattcac ctattaccgt tcaggagaag tcctctactt gtcctgttct gcggactcta
     3901 acccaccggc acagtattct tggacaatta atgggaagtt tcagctatca ggacaaaagc
     3961 tctttatccc ccaaattact acaaagcata gcgggctcta tgcttgctct gttcgtaact
     4021 cagccactgg caaggaaagc tccaaatcca tgacagtaaa agtctctggt aagcggatcc
     4081 cagtatcctt ggcaataggg attggtggtt ctggtggtgg ttctggttgg agccatccgc
     4141 agtttgaaaa aggtggtggt agcggtggtg gttcaggtgg tagtgcatgg tcacaccctc
     4201 agtttgagaa atagaattct gcagtcgacg gtaccgcggg cccgggatcc cgcgactcta
     4261 gataattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     4321 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     4381 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
     4441 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     4501 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     4561 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     4621 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     4681 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     4741 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga
     4801 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     4861 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     4921 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4981 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5041 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5101 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5161 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5221 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5281 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5341 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct
     5401 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5461 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5521 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5581 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5641 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5701 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5761 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     5821 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5881 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5941 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     6001 ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     6061 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     6121 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6181 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct
     6241 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     6301 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     6361 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     6421 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     6481 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     6541 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     6601 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     6661 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6721 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6781 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6841 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6901 agaggccttg acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca
     6961 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7021 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     7141 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7201 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7261 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7321 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7381 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7441 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7501 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7561 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7621 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7681 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7741 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7801 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7861 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7921 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7981 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8041 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     8101 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     8161 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     8221 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8281 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8341 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8401 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8461 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8521 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8581 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8641 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8701 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8761 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8821 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8881 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8941 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     9001 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     9061 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     9121 gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     9181 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     9241 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca
     9301 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact
     9361 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg
     9421 tactacaaac ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
     9481 gtagt
//
