LOCUS       V024758                 7084 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024758
VERSION     V024758
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..2577
                     /label="Kcnn1(NM_001363407)"
                     /note="Kcnn1(NM_001363407)"
                     /gene="Kcnn1"
     CDS             2587..2616
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2623..2646
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2684..2908
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2954..3382
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3396..3725
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3792..4586
                     /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    4760..4881
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4930..4946)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4954..4970
                     /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(4978..5008)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5023..5044
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5332..5917)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6088..6948)
                     /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(6949..7053)
                     /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 accatgagta gccacagcca caatggcagc gtggggcagc ctctgggcag cgggcctgga
     1021 ttcctgggct gggagcctgt ggaccctgag gcaggccgcc ccctgcagcc cacccaaggc
     1081 ccaggcctgc agatggtggc caagggtcag cctgtcaggc tgtcacccgg tggttccagg
     1141 ggccaccccc aggagcagga ggaggaagag gaagaggagg aggaggagga ggacaagaca
     1201 ggctcaggga agcccccaac agtcagccac cgcctgggac accgcagggc cctctttgaa
     1261 aagcgtaaac ggctcagtga ctatgcgctc atctttggca tgtttgggat tgtcgtcatg
     1321 gtgacagaaa cagagctgtc ctggggtgta tacaccaagg agtcactctg ctcttttgct
     1381 ctgaaatgcc tcatcagcct gtccactgtc atcttgcttg gccttgtcat cctgtaccac
     1441 gcccgagaga tccagctgtt cttggtggac aatggtgccg acgactggcg tatcgccatg
     1501 acgtgggagc gcgtgtccct gatctcgctg gagttggtcg tgtgtgccat ccacccggtg
     1561 cccggccact atcgcttcac gtggacggca cgactggcct tctctctggt gccgtcggca
     1621 gccgaggcag acctggatgt gctgctgtcc atccccatgt tcctgcgcct ctacctgctg
     1681 gctcgggtca tgctcctgca cagccgcatc ttcaccgatg catccagccg cagcatcggg
     1741 gcccttaacc gcgtcacctt caacacacgc ttcgtcacca agacgctcat gaccatctgc
     1801 cccggcactg tgttgttggt cttcagcgtc tcctcctgga tcgttgctgc gtggacagtg
     1861 cgcgtgtgtg agaggtacca cgataagcag gaagtgacca gcaacttcct gggagccatg
     1921 tggctcatct ccatcacctt cttgtccatt ggctatggag acatggtgcc gcatacctac
     1981 tgtgggaagg gtgtgtgtct gctcactggc atcatgggag caggctgcac tgcactcgtg
     2041 gtggccgtcg tggctcggaa gttggaactc accaaggctg agaaacacgt gcacaacttc
     2101 atgatggaca cacagctcac caagcgggtc aaaaatgctg ctgcaaacgt tctcagggag
     2161 acatggctca tctacaaaca caccaggctg gtgaagaagc cagaccaagg ccgggttcgg
     2221 aaacaccagc gtaagttcct tcaggccatc catcaggctc agaagctccg aagtgtgaag
     2281 attgaacaag ggaaggtgaa cgatcaggcc aacacgctgg ctgagctggc caaggcacag
     2341 agcatcgcat atgaggtggt gtcagagctg caggcccagc aggaggagtt ggaggcacgc
     2401 ctagccgcct tggagagccg actggatgtc ctgggtgcct ccctgcaggc tctaccaggc
     2461 cttatagccc aagccatatg ccctctacca ccaccctggc ctgggcctgg tcacctggcc
     2521 acagccaccc agagcccaca aagccactgg ctgcccacca tgggatcaga ctgtgggtga
     2581 gatatcgagc agaaactcat ctcagaagag gatctgctcg aggattacaa ggatgacgac
     2641 gataagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2701 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2761 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2821 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2881 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2941 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3001 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3301 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3361 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3661 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3721 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3781 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3841 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3901 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3961 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4021 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4081 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4141 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4201 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4261 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4321 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4381 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4441 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4501 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4561 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4621 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4681 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4741 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4801 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4861 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     5881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6001 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6061 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6121 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6181 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6241 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6301 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6361 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6421 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6481 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6541 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6601 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6661 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6721 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6781 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6841 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6901 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6961 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7021 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7081 cgtc
//
