LOCUS       V025476                 7264 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025476
VERSION     V025476
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..2784
                     /label="CANX(NM_001746)"
                     /note="CANX(NM_001746)"
                     /gene="CANX"
     polyA_signal    2864..3088
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3134..3562
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3576..3905
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3972..4766
                     /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    4940..5061
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5110..5126)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5134..5150
                     /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(5158..5188)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5203..5224
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5512..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 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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat ggaagggaag
     1021 tggttgctgt gtatgttact ggtgcttgga actgctattg ttgaggctca tgatggacat
     1081 gatgatgatg tgattgatat tgaggatgac cttgacgatg tcattgaaga ggtagaagac
     1141 tcaaaaccag ataccactgc tcctccttca tctcccaagg ttacttacaa agctccagtt
     1201 ccaacagggg aagtatattt tgctgattct tttgacagag gaactctgtc agggtggatt
     1261 ttatccaaag ccaagaaaga cgataccgat gatgaaattg ccaaatatga tggaaagtgg
     1321 gaggtagagg aaatgaagga gtcaaagctt ccaggtgata aaggacttgt gttgatgtct
     1381 cgggccaagc atcatgccat ctctgctaaa ctgaacaagc ccttcctgtt tgacaccaag
     1441 cctctcattg ttcagtatga ggttaatttc caaaatggaa tagaatgtgg tggtgcctat
     1501 gtgaaactgc tttctaaaac accagaactc aacctggatc agttccatga caagacccct
     1561 tatacgatta tgtttggtcc agataaatgt ggagaggact ataaactgca cttcatcttc
     1621 cgacacaaaa accccaaaac gggtatctat gaagaaaaac atgctaagag gccagatgca
     1681 gatctgaaga cctattttac tgataagaaa acacatcttt acacactaat cttgaatcca
     1741 gataatagtt ttgaaatact ggttgaccaa tctgtggtga atagtggaaa tctgctcaat
     1801 gacatgactc ctcctgtaaa tccttcacgt gaaattgagg acccagaaga ccggaagccc
     1861 gaggattggg atgaaagacc aaaaatccca gatccagaag ctgtcaagcc agatgactgg
     1921 gatgaagatg cccctgctaa gattccagat gaagaggcca caaaacccga aggctggtta
     1981 gatgatgagc ctgagtacgt acctgatcca gacgcagaga aacctgagga ttgggatgaa
     2041 gacatggatg gagaatggga ggctcctcag attgccaacc ctagatgtga gtcagctcct
     2101 ggatgtggtg tctggcagcg acctgtgatt gacaacccca attataaagg caaatggaag
     2161 cctcctatga ttgacaatcc cagttaccag ggaatctgga aacccaggaa aataccaaat
     2221 ccagatttct ttgaagatct ggaacctttc agaatgactc cttttagtgc tattggtttg
     2281 gagctgtggt ccatgacctc tgacattttt tttgacaact ttatcatttg tgctgatcga
     2341 agaatagttg atgattgggc caatgatgga tggggcctga agaaagctgc tgatggggct
     2401 gctgagccag gcgttgtggg gcagatgatc gaggcagctg aagagcgccc gtggctgtgg
     2461 gtagtctata ttctaactgt agcccttcct gtgttcctgg ttatcctctt ctgctgttct
     2521 ggaaagaaac agaccagtgg tatggagtat aagaaaactg atgcacctca accggatgtg
     2581 aaggaagagg aagaagagaa ggaagaggaa aaggacaagg gagatgagga ggaggaagga
     2641 gaagagaaac ttgaagagaa acagaaaagt gatgctgaag aagatggtgg cactgtcagt
     2701 caagaggagg aagacagaaa acctaaagca gaggaggatg aaattttgaa cagatcacca
     2761 agaaacagaa agccacgaag agagtgagaa ttctgcagat atccagcaca gtggcggccg
     2821 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2881 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2941 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3001 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3061 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3121 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3181 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3241 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3301 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3361 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3421 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3481 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3541 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3601 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3781 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3841 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3901 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3961 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4021 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4081 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4141 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4201 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4261 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4321 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4381 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4441 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4501 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4561 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4621 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4681 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4741 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4801 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4861 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4921 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4981 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5041 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5101 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5161 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5941 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 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 cgtc
//
