LOCUS       V040256                 9165 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040256
VERSION     V040256
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..3845
                     /label="MMS19(NM_022362)"
                     /note="MMS19(NM_022362)"
                     /gene="MMS19"
     CDS             3852..3917
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3941..4529
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4595..4643
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4845..5273
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5287..5616
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5683..6477
                     /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    6653..6774
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6823..6839)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6847..6863
                     /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(6871..6901)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6916..6937
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7225..7813)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7984..8844)
                     /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(8845..8949)
                     /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 cgctgccgcg gctgtggagg
      781 cggcggcgcc tatgggtgcc ctatggggcc tcgtgcacga cttcgtcgtg ggtcagcaag
      841 agggccccgc tgaccaggtg gctgcagatg tgaaatctgg caactataca gtgttacaag
      901 ttgtggaagc ccttgggtcc tctctagaga atccagaacc ccgaactcgg gcacgagcaa
      961 tccagctttt gtcacaggtg ctactccact gtcacacctt gctcctggag aaggaagtgg
     1021 tacacctgat actgttctat gagaaccggc tgaaggacca tcatcttgtg atcccatctg
     1081 tcctgcaggg tttgaaggca cttagcctgt gtgtggccct gcccccaggg ctggctgttt
     1141 ctgtgcttaa agccatcttc caggaagtgc atgtacagtc cctgccacag gtggaccgac
     1201 acacagtcta caatatcatc accaatttta tgcgaacccg ggaagaagag ctaaagagcc
     1261 taggagctga cttcaccttt ggcttcatcc aggtgatgga tggggaaaag gatccccgta
     1321 atcttctggt ggccttccgc atcgtccatg acctcatctc cagggactat agcctgggac
     1381 cctttgtgga ggagttgttt gaagtgacat cctgttattt ccctatcgat tttacccctc
     1441 cacctaatga tccccatggt atccagagag aagacctcat cctgagtctt cgcgctgtgc
     1501 tggcttctac accacgattt gctgagtttc tgctgcccct gttgattgag aaagtggatt
     1561 ctgaggttct gagtgccaag ttggattctc tacagactct gaatgcttgc tgtgctgtgt
     1621 atggacagaa ggaactgaag gacttcctcc ccagcctttg ggcttctatc cgcagagagg
     1681 tgttccagac ggcaagtgag cgggtggagg cagagggcct ggcggccctc cactccctga
     1741 ctgcgtgttt gtctcgctct gtgctgaggg ctgatgctga ggacctcctt gactccttcc
     1801 ttagcaacat tctacaggac tgcaggcacc acctgtgtga accggacatg aaactggtgt
     1861 ggcctagtgc caagctgttg caggcagctg caggtgcatc tgcccgggcc tgtgactctg
     1921 tcaccagcaa tgtactgcct ttactgctgg aacagttcca caagcacagt cagagcagcc
     1981 agcggcggac aatccttgaa atgctcctgg gtttcttgaa gctgcagcag aaatggagct
     2041 atgaagacaa agatcaaagg cctctgaatg gcttcaagga ccagctgtgc tcactggtat
     2101 tcatggctct aacagacccc agcacccagc ttcagcttgt tggcatccgt acactgacag
     2161 tcttgggtgc ccagccagat ctcctatctt atgaggactt ggagctggca gtgggtcacc
     2221 tgtacagact gagcttcctg aaggaggatt cccagagttg cagggtggca gcactggaag
     2281 catcaggaac cctggctgct ctctaccctg tggccttcag cagccacctc gtacccaagc
     2341 tcgctgagga gctgcgtgta ggggagtcaa atttgactaa cggagatgag cccacccaat
     2401 gctcccggca tctgtgctgt ctgcaagcct tgtcagctgt atcaacacat cccagcatcg
     2461 tcaaggagac actgcctctg ctgctgcagc atctctggca agtgaacaga gggaatatgg
     2521 ttgcacaatc cagtgacgtt attgctgtct gtcagagcct cagacagatg gcagaaaaat
     2581 gtcagcagga ccctgagagt tgctggtatt tccaccagac agctatacct tgcctgcttg
     2641 ccttggctgt gcaggcctct atgccagaga aggagccctc agttctgaga aaagtactat
     2701 tggaggatga ggtgttggct gccatggtgt ctgtcattgg cactgctaca acccacctga
     2761 gccctgagtt agctgcccag agtgtgacac acattgtgcc cctcttcttg gatggcaacg
     2821 tgtcctttct gcctgaaaac agcttcccga gcagattcca gccattccag gatggctcct
     2881 cagggcagag gcggctgatt gcactgctta tggcctttgt ctgctccctg cctcgaaatg
     2941 tggaaatccc tcagctgaac caactcatgc gggagctttt ggaactgagc tgctgccaca
     3001 gctgcccctt ttcttccacc gctgctgcca agtgctttgc aggactcctc aacaagcacc
     3061 ctgcagggca gcagctggat gaattcctac agctagctgt ggacaaagtg gaggctggcc
     3121 tgggctctgg gccctgtcgt agtcaggcct tcactcttct tctctgggta acaaaggccc
     3181 tagtgctcag ataccatcct ctcagctcct gccttacagc ccggctcatg ggcctcctga
     3241 gtgacccaga attaggtcca gcagcagctg atggcttctc tctgctcatg tctgactgca
     3301 ctgatgtgct gactcgtgct ggccatgccg aagtgcggat catgttccgc cagcggttct
     3361 tcacagataa tgtgcctgct ttggtccagg gcttccatgc tgctccccaa gatgtgaagc
     3421 caaactactt gaagggtctt tctcatgtac ttaacaggct gcccaagcct gtactcttgc
     3481 cagagctgcc cacgcttctt tccttgctgc tggaggccct gtcctgccct gactgtgtgg
     3541 tgcagctctc caccctcagc tgccttcagc ctcttctact ggaagcaccc caagtcatga
     3601 gtcttcacgt ggacaccctc gtcaccaagt ttctgaacct cagctctagc ccttccatgg
     3661 ctgtccggat cgccgcactg cagtgcatgc atgctctcac tcgcctgccc acccctgtgc
     3721 tgctgccgta caaaccacag gtgattcggg ccttagccaa acccctggat gacaagaaga
     3781 gactggtgcg caaggaagca gtgtcagcca gaggggagtg gtttctgttg gggagccctg
     3841 gcagcggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3901 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3961 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4021 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4081 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4141 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4201 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4261 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4321 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     4381 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4441 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4501 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     4561 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4621 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4681 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4741 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4801 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4861 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4921 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4981 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     5041 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5101 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5161 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     5221 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     5281 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5341 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5401 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5461 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5521 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5581 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5641 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5701 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5761 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5821 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5881 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5941 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     6001 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     6061 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     6121 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     6181 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     6241 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6301 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6361 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6421 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6481 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6541 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6601 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6661 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6721 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6781 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6841 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6901 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6961 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7021 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     7441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7861 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7921 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7981 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8041 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8101 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8161 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8221 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8281 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8341 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8401 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8461 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8521 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8581 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8641 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8701 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8761 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8821 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8881 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8941 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     9001 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     9061 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     9121 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
