LOCUS       V029217                 7432 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029217
VERSION     V029217
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..2088
                     /label="CALCRL(NM_005795)"
                     /note="CALCRL(NM_005795)"
                     /gene="CALCRL"
     CDS             2095..2184
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2208..2796
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2862..2910
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3112..3540
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3554..3883
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3950..4744
                     /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    4920..5041
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5090..5106)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5114..5130
                     /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(5138..5168)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5183..5204
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5492..6080)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6251..7111)
                     /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(7112..7216)
                     /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 ccaccatgga gaaaaagtgt
      721 accctgtatt ttctggttct cttgcctttt tttatgattc ttgttacagc agaattagaa
      781 gagagtcctg aggactcaat tcagttggga gttactagaa ataaaatcat gacagctcaa
      841 tatgaatgtt accaaaagat tatgcaagac cccattcaac aagcagaagg cgtttactgc
      901 aacagaacct gggatggatg gctctgctgg aacgatgttg cagcaggaac tgaatcaatg
      961 cagctctgcc ctgattactt tcaggacttt gatccatcag aaaaagttac aaagatctgt
     1021 gaccaagatg gaaactggtt tagacatcca gcaagcaaca gaacatggac aaattatacc
     1081 cagtgtaatg ttaacaccca cgagaaagtg aagactgcac taaatttgtt ttacctgacc
     1141 ataattggac acggattgtc tattgcatca ctgcttatct cgcttggcat attcttttat
     1201 ttcaagagcc taagttgcca aaggattacc ttacacaaaa atctgttctt ctcatttgtt
     1261 tgtaactctg ttgtaacaat cattcacctc actgcagtgg ccaacaacca ggccttagta
     1321 gccacaaatc ctgttagttg caaagtgtcc cagttcattc atctttacct gatgggctgt
     1381 aattactttt ggatgctctg tgaaggcatt tacctacaca cactcattgt ggtggccgtg
     1441 tttgcagaga agcaacattt aatgtggtat tattttcttg gctggggatt tccactgatt
     1501 cctgcttgta tacatgccat tgctagaagc ttatattaca atgacaattg ctggatcagt
     1561 tctgataccc atctcctcta cattatccat ggcccaattt gtgctgcttt actggtgaat
     1621 ctttttttct tgttaaatat tgtacgcgtt ctcatcacca agttaaaagt tacacaccaa
     1681 gcggaatcca atctgtacat gaaagctgtg agagctactc ttatcttggt gccattgctt
     1741 ggcattgaat ttgtgctgat tccatggcga cctgaaggaa agattgcaga ggaggtatat
     1801 gactacatca tgcacatcct tatgcacttc cagggtcttt tggtctctac cattttctgc
     1861 ttctttaatg gagaggttca agcaattctg agaagaaact ggaatcaata caaaatccaa
     1921 tttggaaaca gcttttccaa ctcagaagct cttcgtagtg cgtcttacac agtgtcaaca
     1981 atcagtgatg gtccaggtta tagtcatgac tgtcctagtg aacacttaaa tggaaaaagc
     2041 atccatgata ttgaaaatgt tctcttaaaa ccagaaaatt tatataatct cgaggaacaa
     2101 aaactaatat cagaggaaga tttggaacag aaactaatca gtgaagaaga tttagagcaa
     2161 aagttaattt ccgaggagga cttatagtga gggcccgata tctcgacaat caacctctgg
     2221 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2281 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2341 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     2401 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     2461 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     2521 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2581 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     2641 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     2701 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     2761 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga
     2821 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     2881 taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     2941 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     3001 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     3061 caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
     3121 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3181 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3241 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     3301 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3361 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3421 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3481 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3541 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3721 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3781 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3841 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3901 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3961 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4021 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4081 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4141 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4201 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4261 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4321 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4381 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4441 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4501 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4561 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4621 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4681 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4741 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4801 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4861 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4921 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4981 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5041 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     5701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7261 atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag
     7321 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7381 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
