LOCUS       V024929                 8017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024929
VERSION     V024929
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             964..3510
                     /label="Kcnn2(NM_001312905)"
                     /note="Kcnn2(NM_001312905)"
                     /gene="Kcnn2"
     CDS             3520..3549
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3556..3579
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    3617..3841
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3887..4315
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4329..4658
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4725..5519
                     /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    5693..5814
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5863..5879)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5887..5903
                     /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(5911..5941)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5956..5977
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6265..6850)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7021..7881)
                     /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(7882..7986)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgttcg ggagaacgtg caaatgtgat gtcatgccca ttgttttggt gcgcccaacc
     1021 aatcggactc gccgcctgga ttctaccgga gccggcatgg gcccttcctc gcaccagcag
     1081 caggagtccc cgctcccgac cataacgcat tgcgcagggt gcaccaccgc ctggtctccc
     1141 tgcagcttta acagctctga catggaaacc ccattgcagt tccagcgcgg cttcttccca
     1201 gagcagccgc cgccaccgcc gcgctcctca cacctgcatt gccagcagca gcaacagagc
     1261 caggacaagc cgtgcgcgcc cttcgcgccc ctcccgcacc ctcaccacca cccgcacctc
     1321 gcgcaccagc agccgggcag cggtggcagc agcccatgcc tccggtgcaa cagctgcgcc
     1381 tcctccggtg ccccggcagc gggggcgggg gcgggggata acctgtccct gctgctccgc
     1441 acctcatcgc ccggcggcgc cttccggacc cgcacctcct cgccactgtc gggctcttcg
     1501 tgctgttgct gctgctcgtc gcgccggggc agccagctca atgtgagcga gctgacgccg
     1561 tccagccatg ccagtgcgct ccggcagcag tacgcgcagc agcccgcgtc cgcctcccag
     1621 taccaccagt gccacagcct gcagcccgcc accagcccca caggcagcct gggcagcctg
     1681 ggctccgggc ccccgctctc gcaccaccac caccacccgc acccggcgca ccaccagcat
     1741 caccaacccc aggcgcgccg cgagagcaac cccttcaccg aaatagccat gagcagctgc
     1801 aggtacaacg ggggcgtcat gcgtccgctc agcaacttga gctcgtcccg ccggaacctg
     1861 caggagatgg actccgaggc tcagcccctg cagccccccg cgtccgtcgt aggaggaggt
     1921 ggtggtgcgt cctccccgtc tgctgccgcc gccgcctcat cctcagcccc ggagatcgtg
     1981 gtgtctaagc cggagcacaa caattctaac aacctggcgc tctacgggac tggcggcgga
     2041 ggcagcaccg gaggcggcgg cggcggcagc gggcatggca gcagcagcgg caccaagtcc
     2101 agcaaaaaga agaaccagaa catcggctac aagctgggcc atcggcgcgc cctgtttgag
     2161 aagcgcaagc ggctcagcga ctatgcgctt atcttcggca tgttcggcat cgtggttatg
     2221 gtcattgaga ccgagctgtc ttggggcgcc tacgacaagg cgtcgctgta ttctttagct
     2281 ctgaaatgcc ttatcagtct ctccacgatc atcctgcttg gtctgatcat cgtgtaccac
     2341 gccagggaaa tacagttatt catggtggac aatggagcag atgactggag aatagccatg
     2401 acttatgagc gtattttctt catctgcttg gaaatactgg tgtgtgctat ccatcccata
     2461 cctgggaatt atacgttcac atggacagcc cggcttgcct tctcctatgc cccctccaca
     2521 accaccgcag atgtggatat tattttatct ataccaatgt tcttaaggct ctatctgatt
     2581 gccagagtca tgctattaca tagcaaactt ttcaccgatg cctcgtctag aagcattggg
     2641 gcacttaata agataaactt caatacgcgt tttgttatga agactttaat gactatctgc
     2701 ccaggaactg tgctcttggt ttttagtatc tcattatgga taattgccgc atggactgtc
     2761 cgagcttgtg aaaggtacca tgatcaacag gatgtcacta gcaacttcct tggagcaatg
     2821 tggttgatat caataacttt tctctccatt ggttatggtg acatggtacc caacacatac
     2881 tgtgggaaag gagtctgctt gcttactgga atcatgggtg caggttgcac agccctggtg
     2941 gtagccgtag tggcaagaaa gctagaactt accaaagcag aaaagcatgt gcacaatttc
     3001 atgatggata ctcagctgac caaaagagtg aaaaacgcag ccgccaatgt actcagggaa
     3061 acgtggttaa tctacaaaaa cacaaagcta gtgaaaaaga tcgaccacgc aaaagtaaga
     3121 aagcatcaac ggaaattctt gcaagctatt catcaattaa gaagtgtgaa gatggaacag
     3181 aggaaactga atgaccaagc aaatacccta gtggatctgg caaagaccca gaatatcatg
     3241 tatgacatga tttccgactt gaatgaaagg agtgaagact ttgagaaaag gatcgtcacc
     3301 ctggaaacaa aactagagac tttgattggt agcattcatg ccctccctgg gctaatcagc
     3361 cagaccatca gacagcagca aagggacttc attgagacac agatggagaa ctacgacaag
     3421 catgtctcct acaatgctga gcgctcccgg tcctcgtcca ggaggcggcg gtcctcctcc
     3481 acagcgccac caacttcatc agagagtagc taggatatcg agcagaaact catctcagaa
     3541 gaggatctgc tcgaggatta caaggatgac gacgataagt agtgagggcc cgtttaaacc
     3601 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3661 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3721 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3781 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3841 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3901 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3961 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4021 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4081 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4141 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4201 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4261 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4321 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4381 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4501 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4561 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4621 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4681 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4741 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4801 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4861 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4921 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4981 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5041 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5101 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5161 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5221 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5281 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5341 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5401 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5461 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5521 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5581 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5641 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5701 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5761 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5821 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5881 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5941 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6001 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6061 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6121 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6181 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6241 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6301 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6361 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6421 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6481 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6541 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6601 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6661 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6721 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6781 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7141 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7981 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
