LOCUS       V037101                 7657 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037101
VERSION     V037101
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"
     CDS             706..2313
                     /label="SRC(NM_198291)"
                     /note="SRC(NM_198291)"
                     /gene="SRC"
     CDS             2320..2409
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2433..3021
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3087..3135
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3337..3765
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3779..4108
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4175..4969
                     /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    5145..5266
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5315..5331)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5339..5355
                     /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(5363..5393)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5408..5429
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5717..6305)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6476..7336)
                     /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(7337..7441)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggg tagcaacaag
      721 agcaagccca aggatgccag ccagcggcgc cgcagcctgg agcccgccga gaacgtgcac
      781 ggcgctggcg ggggcgcttt ccccgcctcg cagaccccca gcaagccagc ctcggccgac
      841 ggccaccgcg gccccagcgc ggccttcgcc cccgcggccg ccgagcccaa gctgttcgga
      901 ggcttcaact cctcggacac cgtcacctcc ccgcagaggg cgggcccgct ggccggtgga
      961 gtgaccacct ttgtggccct ctatgactat gagtctagga cggagacaga cctgtccttc
     1021 aagaaaggcg agcggctcca gattgtcaac aacacagagg gagactggtg gctggcccac
     1081 tcgctcagca caggacagac aggctacatc cccagcaact acgtggcgcc ctccgactcc
     1141 atccaggctg aggagtggta ttttggcaag atcaccagac gggagtcaga gcggttactg
     1201 ctcaatgcag agaacccgag agggaccttc ctcgtgcgag aaagtgagac cacgaaaggt
     1261 gcctactgcc tctcagtgtc tgacttcgac aacgccaagg gcctcaacgt gaagcactac
     1321 aagatccgca agctggacag cggcggcttc tacatcacct cccgcaccca gttcaacagc
     1381 ctgcagcagc tggtggccta ctactccaaa cacgccgatg gcctgtgcca ccgcctcacc
     1441 accgtgtgcc ccacgtccaa gccgcagact cagggcctgg ccaaggatgc ctgggagatc
     1501 cctcgggagt cgctgcggct ggaggtcaag ctgggccagg gctgctttgg cgaggtgtgg
     1561 atggggacct ggaacggtac caccagggtg gccatcaaaa ccctgaagcc tggcacgatg
     1621 tctccagagg ccttcctgca ggaggcccag gtcatgaaga agctgaggca tgagaagctg
     1681 gtgcagttgt atgctgtggt ttcagaggag cccatttaca tcgtcacgga gtacatgagc
     1741 aaggggagtt tgctggactt tctcaagggg gagacaggca agtacctgcg gctgcctcag
     1801 ctggtggaca tggctgctca gatcgcctca ggcatggcgt acgtggagcg gatgaactac
     1861 gtccaccggg accttcgtgc agccaacatc ctggtgggag agaacctggt gtgcaaagtg
     1921 gcggactttg ggctggctcg gctcattgaa gacaatgagt acacggcgcg gcaaggtgcc
     1981 aaattcccca tcaagtggac ggctccagaa gctgccctct atggccgctt caccatcaag
     2041 tcggacgtgt ggtccttcgg gatcctgctg actgagctca ccacaaaggg acgggtgccc
     2101 taccctggga tggtgaaccg cgaggtgctg gaccaggtgg agcggggcta ccggatgccc
     2161 tgcccgccgg agtgtcccga gtccctgcac gacctcatgt gccagtgctg gcggaaggag
     2221 cctgaggagc ggcccacctt cgagtacctg caggccttcc tggaggacta cttcacgtcc
     2281 accgagcccc agtaccagcc cggggagaac ctcctcgagg aacaaaaact aatatcagag
     2341 gaagatttgg aacagaaact aatcagtgaa gaagatttag agcaaaagtt aatttccgag
     2401 gaggacttat agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     2461 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2521 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2581 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2641 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2701 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2761 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2821 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2881 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2941 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3001 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     3061 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     3121 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     3181 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     3241 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     3301 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3361 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3421 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3481 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3541 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3601 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3661 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     3721 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3781 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3841 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3901 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3961 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4021 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4081 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4141 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4201 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4261 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4321 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4381 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4441 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4501 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4561 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4621 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4681 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4741 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4801 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4861 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4921 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4981 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5041 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5101 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5161 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5221 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5281 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5341 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5401 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5461 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5521 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5581 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5641 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5701 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5761 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5821 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5881 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     5941 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6001 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6061 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6121 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6241 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6901 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7141 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7321 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7441 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     7501 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     7561 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     7621 aggcttgacc gacaattgca tgaagaatct gcttagg
//
