LOCUS       V040274                 7665 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040274
VERSION     V040274
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..2345
                     /label="M1AP(NM_138804)"
                     /note="M1AP(NM_138804)"
                     /gene="M1AP"
     CDS             2352..2417
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2441..3029
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3095..3143
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3345..3773
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3787..4116
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4183..4977
                     /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    5153..5274
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5323..5339)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5347..5363
                     /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(5371..5401)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5416..5437
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5725..6313)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6484..7344)
                     /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(7345..7449)
                     /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 ccaccatgca tcctgggcga actactggta
      781 aagggccctc tactcacact cagattgacc agcaacctcc acggcttctc attgtgcaca
      841 ttgctctacc gtcctgggct gacatctgca ccaacctctg tgaggctctg cagaacttct
      901 tctctctagc ctgcagcttg atgggcccca gccgcatgtc cctgttcagt ttatacatgg
      961 tacaagatca gcatgagtgc atcctccctt ttgtgcaagt gaaagggaac tttgctaggt
     1021 tgcagacctg catctcagaa ctccgcatgt tacagagaga agggtgtttc agatcacaag
     1081 gtgcttctct gcggctggca gtagaggatg ggctccagca attcaaacaa tacagcagac
     1141 atgtgaccac aagggcagct ctgacctata cctccctgga gattactatt ctgacttctc
     1201 agcctggaaa agaggtggtc aaacagttgg aggaagggtt gaaagataca gacctagcca
     1261 gagtcaggag gtttcaggtc gttgaggtca caaagggaat cctagagcac gtggactcag
     1321 cgtctcctgt tgaggatacc agcaatgatg agagttctat tctgggaact gacattgacc
     1381 ttcagactat agacaatgat atcgtcagca tggagatttt cttcaaagcc tggctacata
     1441 acagtggaac agaccaagaa caaatccatc ttcttctttc ttcacagtgt ttcagcaaca
     1501 tttccagacc cagagataat ccaatgtgtc tgaaatgtga tctccaagag cgactgctct
     1561 gcccatccct actcgctggc acagctgacg gctccttgag aatggatgac cctaaaggag
     1621 acttcatcac actctaccag atggcttccc agtcatcggc ctctcattac aagctccaag
     1681 tgatcaaggc tttaaaatct agcgggctct gcgagtcatt gacatatgga ctcccgttca
     1741 tcctcagacc tacaagctgt tggcagctgg actgggatga gctggagaca aatcagcaac
     1801 atttccatgc tttgtgtcac agcctgctga aaagggaatg gctgctgtta gccaaggggg
     1861 aaccaccggg cccaggacac agccagagaa ttcctgccag caccttctat gtgatcatgc
     1921 cgtcacactc cctcacactg ctggtaaagg cggtggccac gcgggaactg atgctgccca
     1981 gcaccttccc cctgctacct gaggacccac atgatgatag ccttaagaat gtggagagca
     2041 tgctggacag cctggagctg gagcccacct acaacccctt gcatgttcaa agccacctgt
     2101 actcacacct gagcagcatc tatgccaagc ctcaggggcg gctccaccca cactgggaga
     2161 gccgagctcc gagaaagcat ccctgcaaga ctgggcagtt gcagaccaac cgagctcgag
     2221 ctactgtggc ccccctgcct atgactcctg tcccaggcag agcctccaag atgccagcag
     2281 ccagcaaatc ttcctcagat gccttcttcc tgccttcaga gtgggagaag gatccctcaa
     2341 ggcccggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     2401 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     2461 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2521 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2581 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2641 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2701 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2761 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2821 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     2881 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2941 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3001 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3061 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3121 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3181 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3241 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3301 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3481 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     3541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     3721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4021 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4141 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4681 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4981 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5041 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5101 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5161 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5221 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5281 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5341 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5401 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5461 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5521 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7441 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7501 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7561 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7621 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
