LOCUS       V034632                 7731 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034632
VERSION     V034632
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             887..2500
                     /label="KCNK10(NM_021161)"
                     /note="KCNK10(NM_021161)"
                     /gene="KCNK10"
     CDS             2537..3247
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     polyA_signal    3298..3522
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3568..3996
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4010..4339
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4406..5200
                     /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    5374..5495
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5544..5560)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5568..5584
                     /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(5592..5622)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5637..5658
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5946..6531)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6702..7562)
                     /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(7563..7667)
                     /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 ggctagcaac ttaaggatcc gccaccatgt tttttctcta
      901 cacagacttc tttctttcct tggtggccgt tcccgcagca gcaccggtgt gccagcccaa
      961 gagcgccact aacgggcaac ccccggctcc ggctccgact ccaactccgc gcctgtccat
     1021 ttcctcccga gccacagtgg tagccaggat ggaaggcacc tcccaagggg gcttgcagac
     1081 cgtcatgaag tggaagacgg tggttgccat ctttgtggtt gtggtggtct accttgtcac
     1141 tggcggtctt gtcttccggg cattggagca gccctttgag agcagccaga agaataccat
     1201 cgccttggag aaggcggaat tcctgcggga tcatgtctgt gtgagccccc aggagctgga
     1261 gacgttgatc cagcatgctc ttgatgctga caatgcggga gtcagtccaa taggaaactc
     1321 ttccaacaac agcagccact gggacctcgg cagtgccttt ttctttgctg gaactgtcat
     1381 tacgaccata gggtatggga atattgctcc gagcactgaa ggaggcaaaa tcttttgtat
     1441 tttatatgcc atctttggaa ttccactctt tggtttctta ttggctggaa ttggagacca
     1501 acttggaacc atctttggga aaagcattgc aagagtggag aaggtctttc gaaaaaagca
     1561 agtgagtcag accaagatcc gggtcatctc aaccatcctg ttcatcttgg ccggctgcat
     1621 tgtgtttgtg acgatccctg ctgtcatctt taagtacatc gagggctgga cggccttgga
     1681 gtccatttac tttgtggtgg tcactctgac cacggtgggc tttggtgatt ttgtggcagg
     1741 gggaaacgct ggcatcaatt atcgggagtg gtataagccc ctagtgtggt tttggatcct
     1801 tgttggcctt gcctactttg cagctgtcct cagtatgatc ggagattggc tacgggttct
     1861 gtccaaaaag acaaaagaag aggtgggtga aatcaaggcc catgcggcag agtggaaggc
     1921 caatgtcacg gctgagttcc gggagacacg gcgaaggctc agcgtggaga tccacgataa
     1981 gctgcagcgg gcggccacca tccgcagcat ggagcgccgg cggctgggcc tggaccagcg
     2041 ggcccactca ctggacatgc tgtcccccga gaagcgctct gtctttgctg ccctggacac
     2101 cggccgcttc aaggcctcat cccaggagag catcaacaac cggcccaaca acctgcgcct
     2161 gaaggggccg gagcagctga acaagcatgg gcagggtgcg tccgaggaca acatcatcaa
     2221 caagttcggg tccacctcca gactcaccaa gaggaaaaac aaggacctca aaaagacctt
     2281 gcccgaggac gttcagaaaa tctacaagac cttccggaat tactccctgg acgaggagaa
     2341 gaaagaggag gagacggaaa agatgtgtaa ctcagacaac tccagcacag ccatgctgac
     2401 ggactgtatc cagcagcacg ctgagttgga gaacggaatg atacccacgg acaccaaaga
     2461 ccgggagccg gagaacaact cattacttga agacagaaac gaattcggtg gaggcggatc
     2521 aggaggcggt ggatctatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     2581 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     2641 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2701 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     2761 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     2821 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     2881 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     2941 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     3001 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     3061 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3121 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     3181 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3241 caagtaggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     3301 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3361 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3421 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3481 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3541 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3601 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3661 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3721 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3781 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3841 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3901 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3961 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4021 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4081 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4141 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4201 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4261 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4321 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4381 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4441 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4501 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4561 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4621 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4681 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4741 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4801 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4861 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4921 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4981 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5041 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5101 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5161 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5221 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5281 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5341 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5401 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5461 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5521 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5581 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5641 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5701 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5761 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5821 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5881 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5941 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6001 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6061 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6121 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6181 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6241 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6301 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6361 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6421 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6841 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7381 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7441 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7501 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7621 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7681 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
