LOCUS       V040208                 9975 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040208
VERSION     V040208
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4655
                     /label="SORBS2(NM_001395207)"
                     /note="SORBS2(NM_001395207)"
                     /gene="SORBS2"
     CDS             4662..4727
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4751..5339
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5405..5453
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5655..6083
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6097..6426
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6493..7287
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    7463..7584
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7633..7649)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7657..7673
                     /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(7681..7711)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7726..7747
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8035..8623)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8794..9654)
                     /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(9655..9759)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgaa cacagatagc ggtgggtgtg
      781 ctcgcaaacg tgccgccatg tctgttacgc taacatctgt gaagagagtg caaagttctc
      841 caaacctatt ggctgcaggg cgtgattctc agtcaccaga ctcagcttgg agatcttaca
      901 atgatggcaa tcaggagaca ctgaacggag atgctacata ttcctctctt gcagcaaaag
      961 gttttagaag cgttcgacca aacctacaag ataaaagatc accaactcag agccagataa
     1021 cagtgaatgg aaactcagga ggtgccgtga gtcccatgag ttactatcag aggccgtttt
     1081 ccccctcggc atattctctc ccagcctcac tcaactccag cattgtcatg cagcacggca
     1141 catccctcga ttccacagac acatatcccc agcatgcgca gtctctggat ggcaccacca
     1201 gcagctctat ccccctgtac cgatcctcag aggaagagaa gagagtgaca gtcatcaaag
     1261 ccccgcatta cccagggatc gggcccgtgg atgaatccgg aatccccaca gcaattagaa
     1321 cgacagtcga ccggcccaag gactggtaca agacgatgtt taagcaaatt cacatggtgc
     1381 acaagccgga tgatgacaca gacatgtata atactcctta tacatacaat gcaggtctgt
     1441 acaacccacc ctacagtgct cagtcacacc ctgctgcaaa gacccaaacc tacagacctc
     1501 tttccaaaag ccactccgac aacagcccca atgcctttaa ggatgcgtcc tccccagtgc
     1561 ctcccccaca tgttccacct ccagtcccgc cgcttcgacc aagagatcgg tcttcaacag
     1621 aaaagcatga ctgggatcct ccagacagaa aagtggacac aagaaaattt cggtctgagc
     1681 caaggagtat ttttgaatat gaacctggca agtcatcaat tcttcagcat gaaagaccac
     1741 cccctctccc aacaactccg acacctgttc caagggaacc tggccggaag cctctttcta
     1801 gctcccgact cggagaagtc actggtagcc cctcccctcc gccccggagt ggcgccccca
     1861 caccaagctc gcgcgccccg gctctgtctc ccacccggac tgatcgcata aatccagatg
     1921 acatagattt agaaaatgag ccctggtata aattcttttc agaactggag tttggacgcc
     1981 cgcctcctaa aaagcctctg gactatgttc aagatcattc ttctggtgtt ttcaatgagg
     2041 cctccttgta tcagtcctct atagacagaa gcctggaaag acccatgagt tctgcaagca
     2101 tggccagtga cttcaggaag cggaggaaga gcgagcctgc agtgggtcca ccacggggct
     2161 tgggagatca aagtgcgagc aggactagcc caggccgagt ggacctccca ggatcaagca
     2221 ccactcttac aaagtctttc actagctctt ctccttcttc cccatcaaga gcaaaaggtg
     2281 gggatgatag caaaatatgt ccatcccttt gcagttactc agggctcaac ggcaacccct
     2341 ccagtgagtt agattactgt agcacttaca gacagcactt ggatgtccct cgggactcac
     2401 caagagccat tagtttcaag aacggctggc aaatggcccg gcaaaacgca gaaatctgga
     2461 gcagcacgga agaaaccgtc tctcccaaaa tcaaatcccg gagctgtgac gatctcctaa
     2521 acgacgactg cgatagcttc ccggacccga aagtcaagtc ggaaagcatg ggctccctgt
     2581 tatgtgagga ggactccaag gagagctgcc ccatggcgtg ggggtccccc tacgtcccgg
     2641 aggttcgcag taacggcaga tcaaggatca gacacaggtc agcccgcaac gccccggggt
     2701 tcctgaagat gtacaagaaa atgcaccgca tcaaccgcaa ggacctgatg aactccgagg
     2761 tgatctgctc cgtgaagtcc aggattctgc agtacgagag cgagcagcag cacaaggacc
     2821 tgctgcgcgc ctggagccag tgctccacgg aggaggtgcc ccgggacatg gtgcccacac
     2881 gcatctccga attcgaaaag ctgatccaga agtccaaatc catgccaaat ttaggggatg
     2941 acatgctgtc tcccgtaacc ctagaaccac cgcaaaatgg cctatgtccc aagaggcggt
     3001 tttccattga gtatttgctg gaggaagaaa atcaaagcgg cccccccgct cggggccggc
     3061 gaggctgcca gtctaacgcc ctggtgccca ttcacattga agtcaccagc gatgagcagc
     3121 cccgagcaca cgtggagttt tccgacagcg accaggacgg ggttgtgtcc gaccacagtg
     3181 actacattca cctagagggg tcatccttct gcagtgaaag tgactttgat cacttttcct
     3241 tcacatcctc cgaaagcttt tacggatcca gccaccacca ccaccatcac caccaccacc
     3301 accaccgcca cctcatcagc tcctgcaaag gcaggtgccc ggcctcctac actcgattta
     3361 ccacaatgtt aaaacacgaa agagccagac acgaaaacac cgaggagccc agaaggcaag
     3421 aaatggaccc tggcctttct aaacttgctt ttctagtcag tcctgtgcct ttccggagga
     3481 aaaaaaattc ggctcctaag aaacagactg aaaaggcaaa atgtaaagca tctgtgtttg
     3541 aggctctgga ctctgccctt aaagacatct gtgaccaaat taaagctgaa aagaaaaggg
     3601 ggagcttgcc ggacaacagc atcctgcacc gcctcatcag tgagctgctg ccagatgttc
     3661 ccgagaggaa ctcatccctg agagcgctga ggaggagccc cctgcaccag cctctccacc
     3721 cactgcctcc cgatggtgct attcattgtc caccctacca gaatgactgc gggagaatgc
     3781 cccgcagtgc ctctttccaa gacgtggaca cagccaacag cagctgccac caccaagacc
     3841 gtggcggtgc actccaagac cgtgagtccc ctagaagtta ctcatccact ttgactgaca
     3901 tggggagaag tgcaccaagg gaaagaagag gaactccaga aaaagagaaa ttgcctgcaa
     3961 aagctgttta tgattttaag gctcagacat ctaaggagtt gtcatttaag aaaggagata
     4021 ctgtctacat cctcaggaaa attgatcaaa attggtatga gggagaacac cacgggagag
     4081 tgggcatctt cccgatctca tacgtagaga aactcacacc tcctgagaaa gcacagcctg
     4141 caagaccacc tccgccagcc cagcccggag aaatcggaga agctatagcc aaatacaact
     4201 tcaacgcaga cacaaatgtg gagctgtcac tgagaaaggg agatagagtt attcttctta
     4261 aaagagttga tcaaaactgg tatgaaggta aaatcccagg aaccaacaga caaggcatct
     4321 tccctgtttc ctatgtggag gtcgtcaaga agaacacaaa aggtgctgag gactaccctg
     4381 accctccaat accccacagc tattctagtg ataggattca cagcttgagc tcaaataagc
     4441 cacagcgtcc tgtgtttact catgaaaata ttcaaggtgg gggggaaccg tttcaggctc
     4501 tgtataacta tactcccagg aatgaagatg agctggagct cagagaaagt gatgtcattg
     4561 atgtcatgga aaagtgtgat gacggctggt ttgtggggac ctcaagaaga accaaattct
     4621 ttggtacttt ccccggaaac tacgtcaaga ggctgggatc cgactacaaa gaccatgacg
     4681 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     4741 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4801 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     4861 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     4921 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     4981 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5041 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5101 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5161 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5221 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5281 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5341 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5401 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     5461 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5521 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5581 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     5641 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5701 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5761 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     5821 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5881 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5941 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6001 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     6061 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     6121 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6181 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6241 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6301 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6361 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6421 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     6481 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6541 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6601 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6661 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6721 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6781 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6841 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6901 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     6961 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     7021 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     7081 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     7141 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     7201 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7261 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     7321 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7381 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7441 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     7501 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7561 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     7621 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7681 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7741 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8221 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8341 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8401 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8461 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8521 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8581 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8641 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8701 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8761 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8821 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8881 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8941 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9001 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9061 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9121 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9181 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9241 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9301 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9361 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9421 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9481 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9541 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9601 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9661 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9721 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9781 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     9841 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     9901 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     9961 aagaatctgc ttagg
//
