LOCUS       V033105                 9895 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033105
VERSION     V033105
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             2827..4602
                     /label="Igf2bp2(NM_183029)"
                     /note="Igf2bp2(NM_183029)"
                     /gene="Igf2bp2"
     CDS             4603..4620
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        4683..5182
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5203..5802
                     /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    5816..6404
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6611..7244
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7373..7389)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7397..7413
                     /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(7421..7451)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7466..7487
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7775..8363)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8534..9394)
                     /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(9395..9499)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9547..9681
                     /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 gccaccatga tgaacaagct gtacattggg aacctgagcc ccgccgtcac cgccgacgac
     2881 ctccggcagc tcttcgggga caggaagctg cccctggcgg gacaggtcct actcaagtcc
     2941 ggctacgcct tcgtggacta ccccgaccag aactgggcca tccgcgccat cgagaccctc
     3001 tcgggtaaag tggaattgca tgggaaaatc atggaagttg actactcagt ctctaaaaag
     3061 ctaaggagca gaagaatcca gattcggaac atcccgcctc acctgcagtg ggaggtgttg
     3121 gatgggctgt tggctgaata tgggacagtg gagaacgtgg agcaagtcaa cacagataca
     3181 gaaactgccg ttgtcaacgt cacctatatg acaagagaag aggcaaagct agctattgag
     3241 aagctcagtg ggcatcagtt tgaggactac tccttcaaga tttcctacat ccccgatgaa
     3301 gaggtgagct ctccttcacc ccctcatcgt gcccgggaac aaggccacgg ccccgggagc
     3361 tcttctcagg ccagacagat tgatttcccg ctgcggatcc tggtccccac ccagtttgtt
     3421 ggtgccatca tcggaaagga gggcttgacc ataaagaaca tcactaagca gacccagtcc
     3481 cgggtagaca tccacagaaa ggagaactct ggggctgcag agaagcctgt cacaatccat
     3541 gctaccccag aagggacatc tgaagcatgc cgcatgattc ttgagattat gcaaaaagaa
     3601 gctgatgaga ccaaactggc tgaggaggtt cctctgaaaa tcctggccca caatggcttc
     3661 gttggaagac tgattggcaa agaaggcaga aacctgaaga aaatagaaca tgagacaggg
     3721 accaagataa ccatctcatc cttgcaggat ttgagcattt ataaccccga gagaaccatc
     3781 accgtgaggg gcaccattga agcctgtgcc aatgctgaga tagagattat gaagaagctc
     3841 cgagaggcct ttgagaacga catgctggcc gttaaccaac aagccaatct gatcccaggg
     3901 ctaaacctca gtgcacttgg catcttttcg actggactgt ctgtgcttcc tccaccagca
     3961 gggccccgtg gagttccccc cagtcctccc tatcacccct ttgctaccca ctccggatac
     4021 ttctccagtc tgtaccctca tcaccatttc ggcccattcc cacatcatca ctcctaccca
     4081 gagcaggaga ctgtaagtct cttcatccca acccaggctg tgggtgctat catcgggaag
     4141 aagggggcac acatcaaaca gctcgctcga tttgctggtg cctccatcaa gattgctcca
     4201 gcagaaggtc cagatgtcag tgagaggatg gtcatcatca ctggtcctcc tgaagcccag
     4261 tttaaggctc agggacggat ctttgggaaa ctgaaggaag aaaacttctt taatcccaaa
     4321 gaagaagtga agctggaggc ccacatccga gtcccctcgt cgaccgctgg ccgggtgatt
     4381 ggcaagggcg ggaaaaccgt gaacgagctg cagaacttga caagtgcaga agttatcgtg
     4441 cctcgtgacc aaacgccaga cgagaatgag gaagtgatcg tcagaattat cgggcatttt
     4501 tttgctagcc agactgcaca acgcaagatc agggaaattg tacagcaggt gaagcagcag
     4561 gagcagagat accctcaggg agtcgcccca cagcgcagca agcatcacca tcatcaccat
     4621 taggaattct gcagtcgacg gtaccgcggg cccgggatcc cgcgactcta gataattcta
     4681 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     4741 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     4801 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     4861 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     4921 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     4981 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     5041 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     5101 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     5161 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc
     5221 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     5281 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     5341 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     5401 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     5461 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     5521 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     5581 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5641 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5701 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5761 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca
     5821 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5881 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5941 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6001 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6061 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6121 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6181 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     6241 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6301 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6361 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc
     6421 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     6481 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     6541 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6601 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     6661 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc
     6721 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta
     6781 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg
     6841 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac
     6901 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag
     6961 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga
     7021 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga
     7081 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     7141 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     7201 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     7261 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg
     7321 acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7381 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7441 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7501 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7561 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     8221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8641 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9481 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     9541 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     9601 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     9661 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt
     9721 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt
     9781 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac
     9841 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
