LOCUS       V044942                 7449 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044942
VERSION     V044942
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..2129
                     /label="Sh3bp5(NM_011894)"
                     /note="Sh3bp5(NM_011894)"
                     /gene="Sh3bp5"
     CDS             2136..2201
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2225..2813
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2879..2927
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3129..3557
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3571..3900
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3967..4761
                     /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    4937..5058
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5107..5123)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5131..5147
                     /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(5155..5185)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5200..5221
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5509..6097)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6268..7128)
                     /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(7129..7233)
                     /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 ccaccatgga cacggcgctg aaacggagcc
      781 gctctgacga gccggcggag ctcccgccgc ctgcccggga ggtggaggag aaagaggagg
      841 aggaagagag gatggaacag ggcctggagg aggaggagga agaggtggac ccccggatcc
      901 agggagaact ggagaagtta aatcaatcca cggatgatat caaccgacgg gagactgagc
      961 ttgaggatgc acgccagaag ttccgatctg tcctggtcga ggcaacggtg aaactagacg
     1021 aactggcaaa gaaaatcggc aaagccgtgg aggactcgaa gccctactgg gaggcccgga
     1081 gggtggcaag gcaggctcag ctggaggcgc agaaggccac acaggacttc cagagggcca
     1141 ccgaggtact ccgtgcagcc aaggaaacaa tctctctggc tgagcagcgg ctgttggagg
     1201 atgacaagcg gcagtttgac tctgcttggc aggaaatgct gaaccatgct actcagaggg
     1261 tcatggaggc cgagcagacc aagacaagga gtgagctagt acacaaggag acggcagcca
     1321 ggtacaacgc tgccatgggt cgcatgcgcc agctggagaa gaaactcaag cgagccatca
     1381 acaagtccaa gccttatttt gagctaaagg caaagtacta cgtgcagctt gagcaactga
     1441 agaagacggt ggatgacctt caggccaagc tggccctggc aaagggcgag tacaaggcag
     1501 ccctgaagag cctggagagg atctcagatg agatacacga gcggcggcgt tccaatgcca
     1561 tggggccccg gggctgtggc gtgggggcag agggcagcat cgcctcagtg gagaacctgc
     1621 ctgtaagcaa acctgagcct gacgccattt ctgtggcatc agaagccttt gaagatgaca
     1681 actgcagcaa cttggtgtct gaagatgact cggaaaccca gtctgtgtcc agctttagtt
     1741 cgggaccgac gagtccgtct gaaatgcctg atcagttccc tgcagtggca aggcctggca
     1801 gtctggatct gcctagccct gtgtccctgt cagaatttgg gatgatgttc ccaatactgg
     1861 gtcctcgaag tgaatgcagt ggggcctcct cccctgaatg tgaggttgaa cgaggagaca
     1921 gagcagaagg ggccgagaat aaaatgagtg acaaagccaa caacaaccgt gttctcggca
     1981 gcaccaacgg tggcagtggc aggagcagga gccaaagcag cacgtccctt gagagccagg
     2041 ccttggaaac ccggatgaag cagctctccc tacagtgctc aaaaggaaga gatggaatta
     2101 ttgctgacat aaaaatggtg cagattggcg gatccgacta caaagaccat gacggtgatt
     2161 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     2221 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2281 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2341 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2401 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2461 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2521 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2581 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2641 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2701 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2761 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     2821 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2881 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     2941 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3001 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3061 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     3121 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3181 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3241 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3301 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3361 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3421 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3481 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3541 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3901 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3961 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4021 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4081 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4141 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4201 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4261 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4321 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4381 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4441 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4501 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4561 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4621 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4681 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4741 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4801 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4861 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4921 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4981 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5041 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5101 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5161 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5221 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5281 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5341 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5701 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5941 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6121 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6181 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6241 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6301 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6361 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6421 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6481 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6541 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6601 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6661 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6721 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6781 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6841 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6901 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6961 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7021 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7081 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7141 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7201 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7261 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7321 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7381 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7441 ctgcttagg
//
