LOCUS       V047335                 8187 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047335
VERSION     V047335
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2867
                     /label="CLIC6(NM_001317009)"
                     /note="CLIC6(NM_001317009)"
                     /gene="CLIC6"
     CDS             2874..2939
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2963..3551
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3617..3665
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3867..4295
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4309..4638
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4705..5499
                     /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    5675..5796
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5845..5861)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5869..5885
                     /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(5893..5923)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5938..5959
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6247..6835)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7006..7866)
                     /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(7867..7971)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc cgaggccgcg gagccggagg
      781 gggttgcccc gggtccccag gggccgccgg aggtccccgc gcctctggct gagagacccg
      841 gagagccagg agccgcgggc ggggaggcag aagggccgga ggggagcgag ggcgcagagg
      901 aggcgccgag gggcgccgcc gctgtgaagg aggcaggagg cggcgggcca gacaggggcc
      961 cggaggccga ggcgcggggc acgagggggg cgcacggcga gactgaggcc gaggagggag
     1021 ccccggaggg tgccgaggtg ccccaaggag gggaggagac aagcggcgcg cagcaggtgg
     1081 agggggcgag cccgggacgc ggcgcgcagg gcgagccccg cggggaggct cagagggagc
     1141 ccgaggactc tgcggccccc gagaggcagg aggaggcgga gcagaggcct gaggtcccgg
     1201 aaggtagcgc gtccggggag gcgggggaca gcgtagacgc ggagggcccg ctgggggaca
     1261 acatagaagc ggagggcccg gcgggcgaca gcgtagaggc ggagggccgg gtgggggaca
     1321 gcgtagacgc ggaaggtccg gcgggggaca gcgtagacgc ggagggcccg ctgggggaca
     1381 acatacaagc cgagggcccg gcgggggaca gcgtagacgc ggagggccgg gtgggggaca
     1441 gcgtagacgc ggaaggtccg gcgggggaca gcgtagacgc ggagggccgg gtgggggaca
     1501 gcgtagaggc gggggacccg gcgggggacg gcgtagaagc gggggtcccg gcgggggaca
     1561 gcgtagaagc cgaaggcccg gcgggggaca gcatggacgc cgagggtccg gcaggaaggg
     1621 cgcgccgggt ctcgggtgag ccgcagcaat cgggggacgg cagcctctcg ccccaggccg
     1681 aggcaattga ggtcgcagcc ggggagagtg cggggcgcag ccccggtgag ctcgcctggg
     1741 acgcagcgga ggaggcggag gtcccggggg taaaggggtc cgaagaagcg gcccccgggg
     1801 acgcaagggc agacgctggc gaggacaggg taggggatgg gccacagcag gagccggggg
     1861 aggacgaaga gagacgagag cggagcccgg aggggccaag ggaggaggaa gcagcggggg
     1921 gcgaagagga atcccccgac agcagcccac atggggaggc ctccaggggc gccgcggagc
     1981 ctgaggccca gctcagcaac cacctggccg aggagggccc cgccgagggt agcggcgagg
     2041 ccgcgcgcgt gaacggccgc cgggaggacg gagaggcgtc cgagccccgg gccctggggc
     2101 aggagcacga catcaccctc ttcgtcaagg taaaacttac tgcattaggc tgttctcgca
     2161 ttgctataaa gaaatacctg agagctggtt atgatggtga gagtatcgga aattgcccgt
     2221 tttctcagcg tctctttatg attctctggc tgaaaggcgt tatatttaat gtgaccacag
     2281 tggacctgaa aaggaaaccc gcagacctgc agaacctggc tcccggaaca aaccctcctt
     2341 tcatgacttt tgatggtgaa gtcaagacgg atgtgaataa gatcgaggag ttcttagagg
     2401 agaaattagc tcccccgagg tatcccaagc tggggaccca acatcccgaa tctaattccg
     2461 caggaaatga cgtgtttgcc aaattctcag cgtttataaa aaacacgaag aaggatgcaa
     2521 atgagattca tgaaaagaac ctgctgaagg ccctgaggaa gctggataat tacttaaata
     2581 gccctctgcc tgatgaaata gatgcctaca gcaccgagga tgtcactgtt tctggaagga
     2641 agtttctgga tggggacgag ctgacgctgg ctgactgcaa cctcttaccc aagctccata
     2701 ttattaagat tgtggccaag aagtacagag attttgaatt tccttctgaa atgactggca
     2761 tctggagata cttgaataat gcttatgcta gagatgagtt cacaaatacg tgtccagctg
     2821 atcaagagat tgaacacgca tattcagatg ttgcaaaaag aatgaaagga tccgactaca
     2881 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     2941 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3001 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3061 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3121 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3181 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3241 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3301 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3361 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3421 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3481 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3541 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     3601 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     3661 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3721 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3781 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     3841 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3901 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3961 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4021 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4081 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4141 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4201 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4261 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4321 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4381 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4441 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4501 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4561 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4621 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4681 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4741 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4801 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4861 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4921 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4981 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5041 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5101 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5161 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5221 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5281 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5341 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5401 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5461 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5521 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5581 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5641 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5761 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5821 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5881 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5941 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6001 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6061 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6421 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6661 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7141 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7861 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7981 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     8041 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8101 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8161 gacaattgca tgaagaatct gcttagg
//
