LOCUS       V027269                 6604 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027269
VERSION     V027269
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2040
                     /label="CAMK1(NM_003656)"
                     /note="CAMK1(NM_003656)"
                     /gene="CAMK1"
     CDS             2047..2136
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2171..2395
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2441..2869
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2883..3212
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3279..4073
                     /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    4247..4368
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4417..4433)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4441..4457
                     /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(4465..4495)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4510..4531
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4819..5404)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5575..6435)
                     /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(6436..6540)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgctggggg cagtggaagg ccccaggtgg
      961 aagcaggcgg aggacattag agacatctac gacttccgag atgttctggg cacgggggcc
     1021 ttctcggagg tgatcctggc agaagataag aggacgcaga agctggtggc catcaaatgc
     1081 attgccaagg aggccctgga gggcaaggaa ggcagcatgg agaatgagat tgctgtcctg
     1141 cacaagatca agcaccccaa cattgtagcc ctggatgaca tctatgagag tgggggccac
     1201 ctctacctca tcatgcagct ggtgtcgggt ggggagctct ttgaccgtat tgtggaaaaa
     1261 ggcttctaca cggagcggga cgccagccgc ctcatcttcc aggtgctgga tgctgtgaaa
     1321 tacctgcatg acctgggcat tgtacaccgg gatctcaagc cagagaatct gctgtactac
     1381 agcctggatg aagactccaa aatcatgatc tccgactttg gcctctccaa gatggaggac
     1441 ccgggcagtg tgctctccac cgcctgtgga actccgggat acgtggcccc tgaagtcctg
     1501 gcccagaagc cctacagcaa ggctgtggat tgctggtcca taggtgtcat cgcctacatc
     1561 ttgctctgcg gttaccctcc cttctatgac gagaatgatg ccaaactctt tgaacagatt
     1621 ttgaaggccg agtacgagtt tgactctcct tactgggacg acatctctga ctctgccaaa
     1681 gatttcatcc ggcacttgat ggagaaggac ccagagaaaa gattcacctg tgagcaggcc
     1741 ttgcagcacc catggattgc aggagataca gctctagata agaatatcca ccagtcggtg
     1801 agtgagcaga tcaagaagaa ctttgccaag agcaagtgga agcaagcctt caatgccacg
     1861 gctgtggtgc ggcacatgag gaaactgcag ctgggcacca gccaggaggg gcaggggcag
     1921 acggcgagcc atggggagct gctgacacca gtggctgggg ggccggcagc tggctgttgc
     1981 tgtcgagact gctgcgtgga gccgggcaca gaactgtccc ccacactgcc ccaccagctc
     2041 ctcgaggaac aaaaactaat atcagaggaa gatttggaac agaaactaat cagtgaagaa
     2101 gatttagagc aaaagttaat ttccgaggag gacttatgag ggcccgttta aacccgctga
     2161 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2221 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2281 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2341 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2401 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2461 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2521 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2581 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2641 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2701 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2761 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2821 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2881 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2941 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3061 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3121 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3181 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3241 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3301 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3361 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3421 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3481 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3541 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3601 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3661 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3721 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3781 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3841 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3901 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3961 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4021 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4081 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4141 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4201 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4261 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4321 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4381 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4441 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4501 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4561 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4621 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4681 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4741 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5221 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5281 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5341 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5401 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5461 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5521 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5581 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5641 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5701 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5761 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5821 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5881 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5941 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6001 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6061 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6121 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6181 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6241 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6301 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6361 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6421 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6481 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6541 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6601 tggt
//
