LOCUS       V025183                10017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025183
VERSION     V025183
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..4697
                     /label="RNF123(NM_022064)"
                     /note="RNF123(NM_022064)"
                     /gene="RNF123"
     CDS             4704..4769
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4793..5381
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5447..5495
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5697..6125
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6139..6468
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6535..7329
                     /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    7505..7626
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7675..7691)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7699..7715
                     /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(7723..7753)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7768..7789
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8077..8665)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8836..9696)
                     /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(9697..9801)
                     /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 atccaagggg gccggcatgt
      781 ctttctcccg caagagctat aggctgacct cagatgctga gaaatccagg gtcacaggca
      841 ttgtgcagga gaagctgctg aatgactacc tgaaccgcat cttttcctct tctgaacatg
      901 cacccccagc agccaccagc aggaaacccc tgaacttcca gaacctgcca gaacatttgg
      961 accagttgct acaggtggac aatgaggagg aggaaagcca gggacaggtt gaagggcggc
     1021 ttggcccatc cactgtggtc ctggaccaca caggcggctt tgaggggctt ctcctggtgg
     1081 atgatgacct gctgggggtg attggacaca gcaactttgg caccatccgc tctaccacat
     1141 gcgtgtacaa agggaaatgg ctctacgagg tcctcatctc ctcccagggg ctcatgcaga
     1201 tcggctggtg caccatcagc tgccgcttca accaggagga gggggttgga gatacacaca
     1261 actcctatgc ctatgatggc aaccgcgtgc gcaagtggaa tgtgaccaca acgaattatg
     1321 gcaaggcgtg ggcagcgggg gacatcgtga gctgcctgat tgacctggat gatggcactc
     1381 tgtccttctg cctgaacggt gtatcactgg gcactgcctt tgagaacctg tccaggggcc
     1441 tgggtatggc ctacttccca gccatcagcc tctctttcaa ggagtccgtg gccttcaact
     1501 ttggcagccg tcctctgcgc tacccagtgg caggctaccg gcccctgcag gacccaccga
     1561 gtgctgacct ggtgcgggca cagaggttgc tgggctgctt ccgggcagtg ctgagtgtgg
     1621 agctggaccc tgtggagggg cggctgttgg acaaggagag ctccaagtgg cggttgcggg
     1681 gccagcccac cgtcctcctc acactggccc acatcttcca tcactttgca ccgcttctgc
     1741 gcaaggtgta tctggtggag gctgtgctca tgagcttctt gctgggcatc gtggagaagg
     1801 gcacacccac acaggcacag tccgtggtgc accaggtcct ggacctcttg tggctcttca
     1861 tggaggacta cgaggtacaa gattgcctca agcagttgat gatgtctctg cttcggctgt
     1921 accgattctc acccattgtc ccagacctgg gcctacagat ccattacctg cggctcacta
     1981 tcgccatcct gaggcatgag aagtcccgca agtttctgct tagcaatgtc ctcttcgacg
     2041 tgctccgctc cgtcgtcttc ttttacatca agagccccct gcgtgtggag gaggccggcc
     2101 tgcaggagct cattcccacc acctggtggc cccactgctc cagtagggag ggcaaagaga
     2161 gcacggagat gaaggaggag accgcagagg agcggctgcg gcggcgagcc tacgaacggg
     2221 gctgtcagcg gctcaggaag cgcatcgaag tggtggaaga actacaggtc cagatcctga
     2281 agctgctgct ggacaataaa gatgacaatg ggggtgaagc ttctaggtat atcttcctga
     2341 ccaagtttcg caagtttctg caggagaacg ccagtggccg ggggaacatg cccatgctct
     2401 gcccccctga gtacatggtc tgcttcttac accggctgat ctctgccctg cgctactatt
     2461 gggatgaata caaggcttcc aatcctcatg cttccttcag tgaggaggcc tacatcccgc
     2521 cccaggtctt ctataatggc aaggtggact actttgacct gcagcgcctg gggggcctcc
     2581 tctcgcacct gcggaagacc ctcaaagatg accttgcttc caaagccaac attgtgatcg
     2641 acccactgga gctccagtca accgccatgg atgacctaga tgaggatgag gagccagccc
     2701 cagctatggc ccagcgcccc atgcaggccc tggctgttgg ggggccactg cccctgcccc
     2761 ggcccggctg gctcagttct ccaactttgg gccgagccaa ccgcttcctc agcacagcgg
     2821 ctgtgagcct catgacccca cggcggcctc tgagcacctc ggagaaagtg aaggtccgca
     2881 cgctgagcgt ggagcagagg acccgtgagg acattgaagg cagccactgg aatgagggct
     2941 tgctgctggg gcggcccccc gaggagcctg agcagcccct caccgagaac tcgctgctgg
     3001 aagtcctgga tggggcggtc atgatgtaca acctcagcgt acaccagcag ctgggcaaga
     3061 tggtgggtgt ctccgatgat gtcaatgaat acgctatggc tctgagggac acagaggaca
     3121 agctccgccg gtgccccaag aggaggaagg acatccttgc agagttgacc aagagccaga
     3181 aggttttctc agaaaagctg gaccacctga gccgccgtct tgcctgggtc catgccactg
     3241 tctactccca ggagaagatg ctggacatct actggctgct gcgcgtctgc ctgcggacca
     3301 ttgagcacgg tgatcgcaca gggtctctct ttgccttcat gcccgagttc tacctgagcg
     3361 tggccatcaa cagctacagt gctctcaaga attactttgg tcccgtgcac agcatggagg
     3421 agctcccagg ctatgaagag accctgaccc gcctggctgc cattctcgcc aaacactttg
     3481 ccgacgcacg cattgtgggc actgacatcc gagactcact gatgcaggcc ctggccagct
     3541 acgtgtgcta cccacactcc ctgcgggctg tggagcgaat ccccgaggag cagcgtatcg
     3601 ccatggtgag gaacctcctg gcgccctatg agcagcggcc ctgggcccag accaactgga
     3661 tcctggtgcg gctctggagg ggctgtggct tcgggtaccg ctatacacgg ctgccacatc
     3721 tgctgaaaac caaacttgag gacgccaatt tgcccagcct ccagaagccc tgcccttcca
     3781 ccctgctgca gcagcacatg gcggacctcc tacagcaggg tcctgatgtg gcacccagct
     3841 tcctcaacag cgtcctcaat cagctcaact gggccttctc tgaattcatt ggcatgatcc
     3901 aagagatcca gcaggctgct gagcgcctgg agcggaactt tgtggacagc cggcagctca
     3961 aggtatgtgc cacctgcttt gacctctcgg tcagcctgct gcgtgtcttg gagatgacta
     4021 tcacactggt gcctgagata ttccttgact ggacccggcc tacctctgag atgctgctgc
     4081 ggcgtcttgc acagctgcta aaccaggtgc tgaaccgggt gacagctgag aggaacctgt
     4141 ttgatcgtgt ggtcacccta cggctgcctg gcctagagag cgtggaccac tatcccattc
     4201 tggtggcagt gacgggcatc ctggtgcagc tcctggtgcg tggcccagcc tcagagagag
     4261 agcaagccac atcagtgctc ctggcagatc cctgcttcca gctacgctca atatgctatc
     4321 tcctgggaca gccagagccc ccagcacctg gcactgctct gccagcccct gaccggaagc
     4381 gcttctccct gcagagctat gcggattata tcagtgccga tgagctggcc caagtggaac
     4441 agatgctggc gcacctgacc tctgcatctg cccaggcagc agctgcctcc ctgcccacca
     4501 gtgaggagga cctctgcccc atctgctatg cccaccccat ctctgctgtg ttccagccct
     4561 gtggccacaa gtcctgcaaa gcctgtatca accagcacct gatgaacaac aaggactgct
     4621 tcttctgcaa aaccaccatc gtgtctgtag aggactggga gaagggagcc aatacgagta
     4681 ctacctcctc agctgccgga tccgactaca aagaccatga cggtgattat aaagatcatg
     4741 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     4801 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4861 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4921 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4981 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5041 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5101 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5161 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     5221 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5281 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5341 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     5401 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     5461 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5521 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5581 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5641 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     5701 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5761 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5821 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     5881 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5941 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6001 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6061 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6121 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6181 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     6421 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     6481 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     6541 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     6601 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6661 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     6721 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6781 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6841 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6901 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6961 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     7021 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     7081 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     7141 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     7201 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     7261 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     7321 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     7381 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     7441 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     7501 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     7561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     7621 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     7681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7861 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     8041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     8281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     9841 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     9901 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     9961 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
