LOCUS       V026130                10511 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026130
VERSION     V026130
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"
     regulatory      2820..2829
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2829..2894
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2901..5267
                     /label="CDH9(NM_016279)"
                     /note="CDH9(NM_016279)"
                     /gene="CDH9"
     promoter        5299..5798
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5819..6418
                     /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    6432..7020
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7227..7860
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7989..8005)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8013..8029
                     /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(8037..8067)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8082..8103
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8391..8979)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9150..10010)
                     /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(10011..10115)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10163..10297
                     /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 ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2881 atgacgatga caaggaattc atgaggactt accattatat accattattc atctggacct
     2941 atatgttcca tacagttgac accatcctat tacaagaaaa acctaacagt tatttatcaa
     3001 gcaaaaagat agcgggtctg acaaaagatg acggtaaaat gctacgtcgc accaagcgtg
     3061 gctggatgtg gaatcagttc ttcttattgg aagagtacac aggtactgac acacaatatg
     3121 taggcaagct tcacactgac caagataaag gagatggaaa tttaaaatac atactaacag
     3181 gagatggggc tggcagtcta tttgttatag atgaaaatac aggagacatt catgctgcaa
     3241 agaaactaga cagagaagaa aaatctctgt acattcttcg tgccaaggct atagacagaa
     3301 aaactgggcg gcaggtggaa ccggaatcgg aatttatcat taaaatacat gatatcaatg
     3361 acaatgagcc aaaatttaca aaagacttat acactgccag tgttcctgaa atgtctggag
     3421 tcggtacatc tgttatacaa gtaactgcaa cagatgcaga tgacgccaac tatggaaata
     3481 gtgccaaagt ggtctatagc atattgcaag gacagccata tttttcagtg gacccagaat
     3541 caggcataat aaaaactgca ttaccagaca tgagcagaga aaatagagag cagtaccagg
     3601 ttgttataca ggccaaagac atgggtggcc agatgggagg cctttctgga accaccacag
     3661 tgaacatcac gctgacagat gtcaacaaca accctcctcg atttccccag agtacgtatc
     3721 aatttaattc tcctgagtct gtacctcttg gaactcatct tggaaggata aaagccaatg
     3781 accctgacgt gggggaaaat gcagaaatgg agtatagcat tgctgaagga gatggtgcag
     3841 acatgttcga tgtcatcact gacaaggata cacaggaagg gattataact gtcaaacaga
     3901 atttagattt tgaaaatcaa atgctctata ctttaagagt ggatgcaagt aacactcacc
     3961 ctgatccacg attcttacac ctgggacctt tcaaagatac agctgtggtc aaaatatctg
     4021 tggaagatat agatgagcct cctgtgttca ctaaagtctc ttacttgata gaagtagatg
     4081 aagatgtaaa ggagggcagt atcattggac aggttacagc atacgatcca gatgccagga
     4141 acaatttaat aaagtactct gttgatcggc atactgatat ggaccgtatt tttggtattc
     4201 actcagaaaa tggttctatt ttcactttga aagcccttga ccgggaatca tctccttggc
     4261 ataacatcac tgttacagcc acagaaataa ataacccaaa acaaagtagc cacatccctg
     4321 tcttcatcag aattctagat ataaatgacc atgctccgga atttgccatg tattatgaaa
     4381 catttgtttg tgaaaatgca aaacctgggc agttgattca gactgtcagt gtcatggata
     4441 aggatgaccc tccccgaggt cacaaattct tttttgaacc agtgccagaa tttactctca
     4501 atccgaattt caccattgta gataataaag ataatacagc aggaatcatg actcggaaag
     4561 atggctacag tcgcaacaaa atgagcacct acttattgcc gattttaatc tttgacaacg
     4621 attatccaat tcaaagcagc actggtacac tcactatccg tgtgtgtgcc tgcgataatc
     4681 aaggaaacat gcaatcctgc accgcagaag ccctgatcct ttcagccggc ctgagcacgg
     4741 gagctctcgt tgcgattcta ctctgtgtcc tcatactgct tattttagtc gtgttgtttg
     4801 ctgcattgaa gaggcaaaga aaaaaggaac ctctgataat ttcaaaagac gatgtccggg
     4861 acaacattgt gacctacaac gatgaaggcg gcggggaaga agatacccaa gcttttgaca
     4921 ttggcacatt aaggaatcca gaggcaagag aagacagtaa acttagacgg gatgtaatgc
     4981 ctgaaactat ttttcagata aggaggactg tgcctctgtg ggaaaatatt gatgtacaag
     5041 attttatcca tcgaagatta aaagaaaacg acgcagaccc aagtgcacct ccatatgatt
     5101 cgctggcaac gtatgcctat gaagggaatg attccatagc agattcgctc agttctttgg
     5161 aatctctcac agctgattgt aaccaagatt atgattacct cagtgactgg gggcctcgtt
     5221 tcaaaaaact tgccgatatg tatgggggtg atgatagtga ccgagactaa ggatcccgcg
     5281 actctagata attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     5341 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     5401 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     5461 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     5521 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     5581 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     5641 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     5701 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     5761 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat
     5821 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     5881 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     5941 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     6001 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     6061 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     6121 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     6181 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     6241 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     6301 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6361 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc
     6421 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6481 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6541 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6601 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6661 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6721 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6781 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     6841 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6901 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6961 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     7021 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     7081 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     7141 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     7201 ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga
     7261 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     7321 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     7381 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     7441 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     7501 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     7561 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     7621 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     7681 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7741 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7801 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7861 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     7921 agagtgagag gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg
     7981 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     8041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     8101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     8161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     8221 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     8281 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     8341 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     8401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     8461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     8521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8701 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8761 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8821 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8881 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     8941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     9001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     9061 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     9121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     9181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     9241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     9301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     9481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     9961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    10021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    10081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    10141 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag
    10201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    10261 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
    10321 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
    10381 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta
    10441 aatatgtact acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca
    10501 aacttagtag t
//
