LOCUS       V041722                 9642 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041722
VERSION     V041722
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..4347
                     /label="MECOM(NM_001105077)"
                     /note="MECOM(NM_001105077)"
                     /gene="MECOM"
     polyA_signal    4401..5023
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        5065..5362
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5416..6210
                     /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    6862..6996
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(7002..7020)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7034..7050)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7058..7074
                     /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(7082..7112)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7127..7148
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7436..8024)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8195..9055)
                     /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(9056..9160)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      9187..9615
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgatcttaga cgaattttac
     1021 aatgtgaagt tctgcataga tgccagtcaa ccagatgttg gaagctggct caagtacatt
     1081 agattcgctg gctgttatga tcagcacaac cttgttgcat gccagataaa tgatcagata
     1141 ttctatagag tagttgcaga cattgcgccg ggagaggagc ttctgctgtt catgaagagc
     1201 gaagactatc cccatgaaac tatggcgccg gatatccacg aagaacggca atatcgctgc
     1261 gaagactgtg accagctctt tgaatctaag gctgaactag cagatcacca aaagtttcca
     1321 tgcagtactc ctcactcagc attttcaatg gttgaagagg actttcagca aaaactcgaa
     1381 agcgagaatg atctccaaga gatacacacg atccaggagt gtaaggaatg tgaccaagtt
     1441 tttcctgatt tgcaaagcct ggagaaacac atgctgtcac atactgaaga gagggaatac
     1501 aagtgtgatc agtgtcccaa ggcatttaac tggaagtcca atttaattcg ccaccagatg
     1561 tcacatgaca gtggaaagca ctatgaatgt gaaaactgtg ccaagcaggt tttcacggac
     1621 cctagcaacc ttcagcggca cattcgctct cagcatgtcg gtgcccgggc ccatgcatgc
     1681 ccggagtgtg gcaaaacgtt tgccacttcg tcgggcctca aacaacacaa gcacatccac
     1741 agcagtgtga agccctttat ctgtgaggtc tgccataaat cctatactca gttttcaaac
     1801 ctttgccgtc ataagcgcat gcatgctgat tgcagaaccc aaatcaagtg caaagactgt
     1861 ggacaaatgt tcagcactac gtcttcctta aataaacaca ggaggttttg tgagggcaag
     1921 aaccattttg cggcaggtgg attttttggc caaggcattt cacttcctgg aaccccagct
     1981 atggataaaa cgtccatggt taatatgagt catgccaacc cgggccttgc tgactatttt
     2041 ggcgccaata ggcatcctgc tggtcttacc tttccaacag ctcctggatt ttcttttagc
     2101 ttccctggtc tgtttccttc cggcttgtac cacaggcctc ctttgatacc tgctagttct
     2161 cctgttaaag gactatcaag tactgaacag acaaacaaaa gtcaaagtcc cctcatgaca
     2221 catcctcaga tactgccagc tacacaggat attttgaagg cactatctaa acacccatct
     2281 gtaggggaca ataagccagt ggagctccag cccgagaggt cctctgaaga gaggcccttt
     2341 gagaaaatca gtgaccagtc agagagtagt gaccttgatg atgtcagtac accaagtggc
     2401 agtgacctgg aaacaacctc gggctctgat ctggaaagtg acattgaaag tgataaagag
     2461 aaatttaaag aaaatggtaa aatgttcaaa gacaaagtaa gccctcttca gaatctggct
     2521 tcaataaata ataagaaaga atacagcaat cattccattt tctcaccatc tttagaggag
     2581 cagactgcgg tgtcaggagc tgtgaatgat tctataaagg ctattgcttc tattgctgaa
     2641 aaatactttg gttcaacagg actggtgggg ctgcaagaca aaaaagttgg agctttacct
     2701 tacccttcca tgtttcccct cccatttttt ccagcattct ctcaatcaat gtacccattt
     2761 cctgatagag acttgagatc gttacctttg aaaatggaac cccaatcacc aggtgaagta
     2821 aagaaactgc agaagggcag ctctgagtcc ccctttgatc tcaccactaa gcgaaaggat
     2881 gagaagccct tgactccagt cccctccaag cctccagtga cacctgccac aagccaagac
     2941 cagcccctgg atctaagtat gggcagtagg agtagagcca gtgggacaaa gctgactgag
     3001 cctcgaaaaa accacgtgtt tgggggaaaa aaaggaagca acgtcgaatc aagacctgct
     3061 tcagatggtt ccttgcagca tgcaagaccc actcctttct ttatggaccc tatttacaga
     3121 gtagagaaaa gaaaactaac tgacccactt gaagctttaa aagagaaata cttgaggcct
     3181 tctccaggat tcttgtttca cccacaattc caactgcctg atcagagaac ttggatgtca
     3241 gctattgaaa acatggcaga aaagctagag agcttcagtg ccctgaaacc tgaggccagt
     3301 gagctcttac agtcagtgcc ctctatgttc aacttcaggg cgcctcccaa tgccctgcca
     3361 gagaaccttc tgcggaaggg aaaggagcgc tatacctgca gatactgtgg caagattttt
     3421 ccaaggtctg caaacctaac acggcacttg agaacccaca caggagagca gccttacaga
     3481 tgcaaatact gtgacagatc atttagcata tcttctaact tgcaaaggca tgttcgcaac
     3541 atccacaata aagagaagcc atttaagtgt cacttatgtg ataggtgttt tggtcaacaa
     3601 accaatttag acagacacct aaagaaacat gagaatggga acatgtccgg tacagcaaca
     3661 tcgtcgcctc attctgaact ggaaagtaca ggtgcgattc tggatgacaa agaagatgct
     3721 tacttcacag aaattcgaaa tttcattggg aacagcaacc atggcagcca atctcccagg
     3781 aatgtggagg agagaatgaa tggcagtcat tttaaagatg aaaaggcttt ggtgaccagt
     3841 caaaattcag acttgctgga tgatgaagaa gttgaagatg aggtgttgtt agatgaggag
     3901 gatgaagaca atgatattac tggaaaaaca ggaaaggaac cagtgacaag taatttacat
     3961 gaaggaaacc ctgaggatga ctatgaagaa accagtgccc tggagatgag ttgcaagaca
     4021 tccccagtga ggtataaaga ggaagaatat aaaagtggac tttctgctct agatcatata
     4081 aggcacttca cagatagcct caaaatgagg aaaatggaag ataatcaata ttctgaagct
     4141 gagctgtctt cttttagtac ttcccatgtg ccagaggaac ttaagcagcc gttacacaga
     4201 aagtccaaat cgcaggcata tgctatgatg ctgtcactgt ctgacaagga gtccctccat
     4261 tctacatccc acagttcttc caacgtgtgg cacagtatgg ccagggctgc ggcggaatcc
     4321 agtgctatcc agtccataag ccacgtatag gaattcatcg atagatctga tatcggtacc
     4381 agtcgactct agaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     4441 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     4501 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     4561 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     4621 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     4681 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     4741 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     4801 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     4861 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac
     4921 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg
     4981 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt
     5041 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5101 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5161 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5221 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5281 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5341 agtagtgagg aggctttttt ggaggaattg atcagcttgg gatctgatca agagacagga
     5401 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5461 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5521 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5581 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5641 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5701 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5761 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5821 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5881 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5941 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6001 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6061 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6121 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6181 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6241 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6301 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6361 tgctggagtt cttcgcccac cccgggctcg atcccctcgc gagttggttc agctgctgcc
     6421 tgaggctgga cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc atccaggaaa
     6481 ccagcagcgg ctatccgcgc atccatgccc ccgaactgca ggagtgggga ggcacgatgg
     6541 ccgctttggt cgacccggac gggacgctcc tgcgcctgat acagaacgaa ttgcttgcag
     6601 gcatctcatg agtgtgtctt cccgttttcc gcctgaggtc actgcgtgga tggagcgctg
     6661 gcgcctgctg cgcgacggcg agctgctcac cacccactcg ccaagctgga accgtaaaaa
     6721 ggccgcgttg ctggcgtttt tccataggct ccgccgatca taatcagcca taccacattt
     6781 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     6841 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     6901 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6961 tccaaactca tcaatgtatc ttatcatgtc tggatcaatt ccctatagtg agtcgtatta
     7021 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7081 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7141 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7201 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7261 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7321 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7381 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7441 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7501 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7561 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7621 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7681 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7741 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7801 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7861 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7921 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7981 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8041 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8101 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8161 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8221 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8281 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8341 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8401 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8461 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8521 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8581 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8641 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8701 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8761 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8821 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8881 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8941 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9001 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9061 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9121 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9181 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     9241 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     9301 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     9361 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     9421 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     9481 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     9541 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     9601 aatttaacgc gaattttaac aaaatattaa cgcttacaat tt
//
