LOCUS       V030415                 7354 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030415
VERSION     V030415
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..2010
                     /label="AP2M1(NM_004068)"
                     /note="AP2M1(NM_004068)"
                     /gene="AP2M1"
     CDS             2017..2106
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2130..2718
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2784..2832
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3034..3462
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3476..3805
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3872..4666
                     /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    4842..4963
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5012..5028)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5036..5052
                     /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(5060..5090)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5105..5126
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5414..6002)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6173..7033)
                     /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(7034..7138)
                     /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 ccaccatgat tggaggctta
      721 ttcatctata atcacaaggg ggaggtgctc atctcccgag tctaccgaga tgacatcggg
      781 aggaacgcag tggatgcctt tcgggtcaat gttatccatg cccggcagca ggtgcgcagc
      841 cccgtcacca acattgctcg caccagcttc ttccacgtta agcggtccaa catttggctg
      901 gcagcagtca ccaagcagaa tgtcaacgct gccatggtct tcgaattcct ctataagatg
      961 tgtgacgtga tggctgccta ctttggcaag atcagcgagg aaaacatcaa gaacaatttt
     1021 gtgctcatat atgagctgct ggatgagatt ctagactttg gctacccaca gaattccgag
     1081 acaggcgcgc tgaaaacctt catcacgcag cagggcatca agagtcagca tcagacaaaa
     1141 gaagagcagt cacagatcac cagccaggta actgggcaga ttggctggcg gcgagagggt
     1201 atcaagtatc gtcggaatga gctcttcctg gatgtgctgg agagtgtgaa cctgctcatg
     1261 tccccacaag ggcaggtgct gagtgcccat gtgtcgggcc gggtggtgat gaagagctac
     1321 ctgagtggca tgcctgaatg caagtttggg atgaatgaca agattgttat tgaaaagcag
     1381 ggcaaaggca cagctgatga aacaagcaag agcgggaagc aatcaattgc cattgatgac
     1441 tgcaccttcc accagtgtgt gcgactcagc aagtttgact ctgaacgcag catcagcttt
     1501 atcccgccag atggagagtt tgagcttatg aggtatcgca caaccaagga catcatcctt
     1561 cccttccggg tgatcccgct agtgcgagaa gtgggacgca ccaaactgga ggtcaaggtg
     1621 gtcatcaagt ccaactttaa accctcactg ctggctcaga agatcgaggt gaggatccca
     1681 accccactga acacaagcgg ggtgcaggtg atctgcatga aggggaaggc caagtacaag
     1741 gccagcgaga atgccatcgt gtggaagatc aagcgcatgg caggcatgaa ggaatcgcag
     1801 atcagcgcag agattgagct tctgcctacc aacgacaaga agaaatgggc tcgacccccc
     1861 atttccatga actttgaggt gccattcgcg ccctctggcc tcaaggtgcg ctacttgaag
     1921 gtgtttgaac cgaagctgaa ctacagcgac catgatgtca tcaaatgggt gcgctacatt
     1981 ggccgcagtg gcatttatga aactcgctgc ctcgaggaac aaaaactaat atcagaggaa
     2041 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag
     2101 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     2161 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2221 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2281 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2341 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2401 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2461 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2521 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2581 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2641 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2701 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     2761 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     2821 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     2881 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     2941 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     3001 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3061 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3121 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3181 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3241 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3301 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3361 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     3421 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3481 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3541 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3601 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3661 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3721 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3781 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3841 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3901 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3961 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4021 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4081 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4141 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4201 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4261 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4321 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4381 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4441 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4501 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4561 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4621 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4681 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4741 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4801 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4861 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4921 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4981 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5041 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5101 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5161 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5221 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5281 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5341 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5401 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5461 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5521 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5581 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     5641 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5701 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5761 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5821 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5881 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5941 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6001 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6061 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6121 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6181 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6241 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6301 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6361 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6421 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6481 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6781 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7141 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7201 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7261 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7321 cttgaccgac aattgcatga agaatctgct tagg
//
