LOCUS       V027911                 7523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027911
VERSION     V027911
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3013
                     /label="CALCOCO1(NM_020898)"
                     /note="CALCOCO1(NM_020898)"
                     /gene="CALCOCO1"
     CDS             3020..3085
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3123..3347
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3393..3821
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3835..4164
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4231..5025
                     /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    5199..5320
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5369..5385)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5393..5409
                     /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(5417..5447)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5462..5483
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5771..6356)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6527..7387)
                     /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(7388..7492)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggaagaat caccactaag
      961 ccgggcacca tcccgtggtg gagtcaactt tctcaatgta gcccggacct acatccccaa
     1021 caccaaggtg gaatgtcact acacccttcc cccaggcacc atgcccagtg ccagtgactg
     1081 gattggcatc ttcaaggtgg aggctgcctg tgttcgggat taccacacat ttgtgtggtc
     1141 ttccgtgcct gaaagtacaa ctgatggttc ccccattcac accagtgtcc agttccaagc
     1201 cagctacctg cccaaaccag gagctcagct ctaccagttc cgatatgtga accgccaggg
     1261 ccaggtgtgt gggcagagcc cccctttcca gttccgagag ccaaggccca tggatgaact
     1321 ggtgaccctg gaggaggctg atgggggctc tgacatcctg ctggttgtcc ccaaggcaac
     1381 tgtgttacag aaccagctcg atgagagcca gcaagaacgg aatgacctga tgcagctgaa
     1441 gctacagctg gagggacagg tgacagagct gaggagccga gtgcaggagc tcgagagggc
     1501 tctggcaact gccaggcagg agcacacgga gctgatggaa cagtacaagg ggatttcccg
     1561 gtcccatggg gagatcacag aagagaggga catcctgagc cggcaacagg gagaccatgt
     1621 ggcacgcatc ctggagctag aggatgacat ccagaccatc agtgagaaag tgctgacgaa
     1681 ggaagtggag ctggacaggc ttagagacac agtgaaggcc ctgactcggg aacaagagaa
     1741 gctccttggg caactgaaag aagtacaagc agacaaggag caaagtgagg ctgagctcca
     1801 agtggcacaa caggagaacc atcacttaaa tttggacctg aaggaggcga agagctggca
     1861 agaggagcag agtgctcagg ctcagcgact gaaagacaag gtggcccaga tgaaggacac
     1921 cctaggccag gcccagcagc gggtggccga gctggagccc ttgaaggagc agcttcgagg
     1981 ggcccaggag cttgcagcct caagccagca gaaagccacc cttcttgggg aggagttggc
     2041 cagtgcagca gcagccaggg accgcaccat agccgaacta caccgcagcc gcctggaagt
     2101 ggctgaagtt aacggcaggc tggctgagct cggtttgcac ttgaaggaag aaaaatgcca
     2161 atggagcaag gagcgggcag ggctgctgca gagtgtggag gcagagaagg acaagatcct
     2221 gaagctgagt gcagagatac ttcgattgga gaaggcagtt caggaggaga ggacccaaaa
     2281 ccaagtgttc aagactgagc tggcccggga gaaggattct agcctggtac agttgtcaga
     2341 aagtaagcgg gagctgacag agctgcggtc agccctgcgt gtgctccaga aggaaaagga
     2401 gcagttacag gaggagaaac aggaattgct agagtacatg agaaagctag aggcccgcct
     2461 ggagaaggtg gcagatgaga agtggaatga ggatgccacc acagaggatg aggaggccgc
     2521 tgtggggctg agctgcccgg cagctctgac agactcagag gacgagtccc cagaagacat
     2581 gaggctccca ccctatggcc tttgtgagcg tggagaccca ggctcctctc ctgctgggcc
     2641 tcgagaggct tctccccttg ttgtcatcag ccagccggct cccatttctc ctcacctctc
     2701 tgggccagct gaggacagta gctctgactc ggaggctgaa gatgagaagt cagtcctgat
     2761 ggcagctgtg cagagtgggg gtgaggaggc caacttactg cttcctgaac tgggcagtgc
     2821 cttctatgac atggccagtg gctttacagt gggtaccctg tcagaaacca gcactggggg
     2881 ccctgccacc cccacatgga aggagtgtcc tatctgtaag gagcgctttc ctgctgagag
     2941 tgacaaggat gccctggagg accacatgga tggacacttc tttttcagca cccaggaccc
     3001 cttcaccttt gagctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     3061 cgactacaag gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc
     3121 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3181 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3241 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3301 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3361 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3421 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3481 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3541 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3601 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3661 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3721 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3781 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3841 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4081 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4141 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4201 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4261 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4321 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4381 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4441 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4501 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4561 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4621 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4681 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4741 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4801 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4861 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4921 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4981 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5041 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5101 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5161 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5341 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5401 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5461 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5521 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5581 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5641 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5701 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5761 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5821 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5881 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5941 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6001 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6061 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6121 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6181 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6241 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6301 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6361 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6421 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6481 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6541 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6601 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6661 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6721 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6781 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6841 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6901 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6961 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7021 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7081 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7141 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7201 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7261 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7321 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7381 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7441 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7501 cccgaaaagt gccacctgac gtc
//
