LOCUS       V039805                 8079 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039805
VERSION     V039805
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..2759
                     /label="Shcbp1(NM_011369)"
                     /note="Shcbp1(NM_011369)"
                     /gene="Shcbp1"
     CDS             2766..2831
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2855..3443
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3509..3557
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3759..4187
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4201..4530
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4597..5391
                     /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    5567..5688
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5737..5753)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5761..5777
                     /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(5785..5815)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5830..5851
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6139..6727)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6898..7758)
                     /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(7759..7863)
                     /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 ccaccatggc tgatgatttg cgggctggtg
      781 gagttctgga acctatagct atggttccac cgagacctga cttggcggcg gagaaggaac
      841 cggcgtcctg gaaggaaggt ttattcttgg atgcagatcc atgcagtgat caaggctatc
      901 atgctaatcc aggtgctact gtaaaaactc tcataccaga aggaaaaact ccttttccac
      961 gaattatcca aacaaatgaa cttctgtttt atgaacgatt cagagcctat caagattaca
     1021 ttttagctga ctgtaaggcc tctgaggtaa aggaattcac agtcagcttc ttggaaaagg
     1081 tccttgaacc atctggatgg tgggcagtct ggcacactaa tgtgtttgag gtgttggttg
     1141 aggttacaaa tgtggacttt ccatccctga aggcggtcgt aaggcttgca gagccatgca
     1201 tctatgaatc taaattgagc acgtttacct tggccaatgt gaaggagctt ttggacctga
     1261 aggagtttca tctgcctctg caggagttgt gggtggtatc agatgactca catgaattcc
     1321 accagatggc acttgcaatt gagcacgtca gatttttcta caaacacatc tggaggagtt
     1381 gggatgagga agaggaggat gagtatgact attttgtcag atgtgttgaa cctcgactga
     1441 gattgtatta tgacatactt gaagatcgag ttccctcggg acttattgtt gactaccaca
     1501 atctgttgtc tcaatgtgaa gagagttaca ggaaattttt aaatctgaga agcagtttgt
     1561 ccaattgtaa ttctgattct gagcaggaaa atatctccat ggtggaaggg ttaaatttgt
     1621 attcagaaat tgaacagttg aaacaaaagc taaagctcat tgagaatcct ttgttaagat
     1681 atgtttttgg ttatcagaag aactctaata tccaaggaaa gggtactcgt caaaatggcc
     1741 agaaggtcat ccatgtggtt tcctccacca tgaagacagg tctacttcgg tctctattca
     1801 aggacaggtt ttgtgaggag tcttgcaaag aagaaacaga aattaagttc catagtgatc
     1861 tgttgtctgg tataaatgcc tgctatgatg gtgacactgt cattatttgt cctggccatt
     1921 atgtagttca tggcacctgt tccatagctg actccattga gttggaagga tatggcctac
     1981 cagatgacat tgtcatagaa aagaggggca aaggagatac ttttgtggat tgcacgggta
     2041 tggatgttaa aatttcaggc ataaaattta ttcagcatga ttctgtggaa ggaatcttaa
     2101 tcattcacca tggcaagacc acactggaaa actgtgtact acaatgtgaa accacaggag
     2161 tcacagtgcg cacatcagca gaacttttca tgaaaaactc agatgtatat ggtgccaagg
     2221 gtgctggtat agaaatatat cctggaagta aatgtaccct gactgacaat ggaatccatc
     2281 actgcaagga aggaattctc attaaggact tccttgatga acattatgat attcccaaaa
     2341 tatcgatgat aaataacgtc atacacaata atgaaggtta tggtgttgtt ttggtgaagc
     2401 ctacaatttt ctgtgatcta caggaaaata cacaagatga aattaatgac aatatggttc
     2461 agaaaaataa agaggcagat gtcactgaag gattagatct ggaagaaatg cttcagtgtg
     2521 tggctagcaa aatggagcct tatgccacag ctgactttaa tgaacaagct aagggaaact
     2581 gtgaaattat aaatgaacta cttgctattt ccatgcaaaa aggccggatg aagaaaagac
     2641 tgagtgaact tgggattaca caagctgatg acaacataat gtcacaggag atgtttattg
     2701 aaattatggg gaaccagttt aagtggaatg gcaaagggag ttttggcaca tttctttacg
     2761 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     2821 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     2881 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2941 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3001 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3061 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3121 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3181 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3241 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3301 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3361 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3421 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3481 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3541 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3601 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3661 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3721 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3781 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3841 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3901 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3961 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4021 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4081 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4141 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4201 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4261 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4321 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4381 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4441 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4501 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4561 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4621 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4681 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4741 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4801 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4861 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4921 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4981 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5041 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5101 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5161 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5221 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5281 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5341 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5401 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5461 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5521 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5581 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5641 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5701 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5761 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5821 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5881 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5941 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6001 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6061 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6301 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6541 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6661 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6721 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6781 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6841 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6901 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6961 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7021 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7081 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7141 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7201 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7261 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7321 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7381 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7441 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7501 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7561 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7621 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7681 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7741 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7801 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7861 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7921 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7981 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8041 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
