LOCUS       V022606                 9071 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022606
VERSION     V022606
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..4561
                     /label="EGF(NM_001963)"
                     /note="EGF(NM_001963)"
                     /gene="EGF"
     CDS             4568..4633
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4671..4895
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4941..5369
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5383..5712
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5779..6573
                     /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    6747..6868
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6917..6933)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6941..6957
                     /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(6965..6995)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7010..7031
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7319..7904)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8075..8935)
                     /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(8936..9040)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgctgctca ctcttatcat
      961 tctgttgcca gtagtttcaa aatttagttt tgttagtctc tcagcaccgc agcactggag
     1021 ctgtcctgaa ggtactctcg caggaaatgg gaattctact tgtgtgggtc ctgcaccctt
     1081 cttaattttc tcccatggaa atagtatctt taggattgac acagaaggaa ccaattatga
     1141 gcaattggtg gtggatgctg gtgtctcagt gatcatggat tttcattata atgagaaaag
     1201 aatctattgg gtggatttag aaagacaact tttgcaaaga gtttttctga atgggtcaag
     1261 gcaagagaga gtatgtaata tagagaaaaa tgtttctgga atggcaataa attggataaa
     1321 tgaagaagtt atttggtcaa atcaacagga aggaatcatt acagtaacag atatgaaagg
     1381 aaataattcc cacattcttt taagtgcttt aaaatatcct gcaaatgtag cagttgatcc
     1441 agtagaaagg tttatatttt ggtcttcaga ggtggctgga agcctttata gagcagatct
     1501 cgatggtgtg ggagtgaagg ctctgttgga gacatcagag aaaataacag ctgtgtcatt
     1561 ggatgtgctt gataagcggc tgttttggat tcagtacaac agagaaggaa gcaattctct
     1621 tatttgctcc tgtgattatg atggaggttc tgtccacatt agtaaacatc caacacagca
     1681 taatttgttt gcaatgtccc tttttggtga ccgtatcttc tattcaacat ggaaaatgaa
     1741 gacaatttgg atagccaaca aacacactgg aaaggacatg gttagaatta acctccattc
     1801 atcatttgta ccacttggtg aactgaaagt agtgcatcca cttgcacaac ccaaggcaga
     1861 agatgacact tgggagcctg agcagaaact ttgcaaattg aggaaaggaa actgcagcag
     1921 cactgtgtgt gggcaagacc tccagtcaca cttgtgcatg tgtgcagagg gatacgccct
     1981 aagtcgagac cggaagtact gtgaagatgt taatgaatgt gctttttgga atcatggctg
     2041 tactcttggg tgtaaaaaca cccctggatc ctattactgc acgtgccctg taggatttgt
     2101 tctgcttcct gatgggaaac gatgtcatca acttgtttcc tgtccacgca atgtgtctga
     2161 atgcagccat gactgtgttc tgacatcaga aggtccctta tgtttctgtc ctgaaggctc
     2221 agtgcttgag agagatggga aaacatgtag cggttgttcc tcacccgata atggtggatg
     2281 tagccagctc tgcgttcctc ttagcccagt atcctgggaa tgtgattgct ttcctgggta
     2341 tgacctacaa ctggatgaaa aaagctgtgc agcttcagga ccacaaccat ttttgctgtt
     2401 tgccaattct caagatattc gacacatgca ttttgatgga acagactatg gaactctgct
     2461 cagccagcag atgggaatgg tttatgccct agatcatgac cctgtggaaa ataagatata
     2521 ctttgcccat acagccctga agtggataga gagagctaat atggatggtt cccagcgaga
     2581 aaggcttatt gaggaaggag tagatgtgcc agaaggtctt gctgtggact ggattggccg
     2641 tagattctat tggacagaca gagggaaatc tctgattgga aggagtgatt taaatgggaa
     2701 acgttccaaa ataatcacta aggagaacat ctctcaacca cgaggaattg ctgttcatcc
     2761 aatggccaag agattattct ggactgatac agggattaat ccacgaattg aaagttcttc
     2821 cctccaaggc cttggccgtc tggttatagc cagctctgat ctaatctggc ccagtggaat
     2881 aacgattgac ttcttaactg acaagttgta ctggtgcgat gccaagcagt ctgtgattga
     2941 aatggccaat ctggatggtt caaaacgccg aagacttacc cagaatgatg taggtcaccc
     3001 atttgctgta gcagtgtttg aggattatgt gtggttctca gattgggcta tgccatcagt
     3061 aatgagagta aacaagagga ctggcaaaga tagagtacgt ctccaaggca gcatgctgaa
     3121 gccctcatca ctggttgtgg ttcatccatt ggcaaaacca ggagcagatc cctgcttata
     3181 tcaaaacgga ggctgtgaac atatttgcaa aaagaggctt ggaactgctt ggtgttcgtg
     3241 tcgtgaaggt tttatgaaag cctcagatgg gaaaacgtgt ctggctctgg atggtcatca
     3301 gctgttggca ggtggtgaag ttgatctaaa gaaccaagta acaccattgg acatcttgtc
     3361 caagactaga gtgtcagaag ataacattac agaatctcaa cacatgctag tggctgaaat
     3421 catggtgtca gatcaagatg actgtgctcc tgtgggatgc agcatgtatg ctcggtgtat
     3481 ttcagaggga gaggatgcca catgtcagtg tttgaaagga tttgctgggg atggaaaact
     3541 atgttctgat atagatgaat gtgagatggg tgtcccagtg tgcccccctg cctcctccaa
     3601 gtgcatcaac accgaaggtg gttatgtctg ccggtgctca gaaggctacc aaggagatgg
     3661 gattcactgt cttgatattg atgagtgcca actgggggag cacagctgtg gagagaatgc
     3721 cagctgcaca aatacagagg gaggctatac ctgcatgtgt gctggacgcc tgtctgaacc
     3781 aggactgatt tgccctgact ctactccacc ccctcacctc agggaagatg accaccacta
     3841 ttccgtaaga aatagtgact ctgaatgtcc cctgtcccac gatgggtact gcctccatga
     3901 tggtgtgtgc atgtatattg aagcattgga caagtatgca tgcaactgtg ttgttggcta
     3961 catcggggag cgatgtcagt accgagacct gaagtggtgg gaactgcgcc acgctggcca
     4021 cgggcagcag cagaaggtca tcgtggtggc tgtctgcgtg gtggtgcttg tcatgctgct
     4081 cctcctgagc ctgtgggggg cccactacta caggactcag aagctgctat cgaaaaaccc
     4141 aaagaatcct tatgaggagt cgagcagaga tgtgaggagt cgcaggcctg ctgacactga
     4201 ggatgggatg tcctcttgcc ctcaaccttg gtttgtggtt ataaaagaac accaagacct
     4261 caagaatggg ggtcaaccag tggctggtga ggatggccag gcagcagatg ggtcaatgca
     4321 accaacttca tggaggcagg agccccagtt atgtggaatg ggcacagagc aaggctgctg
     4381 gattccagta tccagtgata agggctcctg tccccaggta atggagcgaa gctttcatat
     4441 gccctcctat gggacacaga cccttgaagg gggtgtcgag aagccccatt ctctcctatc
     4501 agctaaccca ttatggcaac aaagggccct ggacccacca caccaaatgg agctgactca
     4561 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     4621 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     4681 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4741 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4801 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4861 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4921 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4981 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5041 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5101 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5161 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5221 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5281 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5341 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5401 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5461 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5521 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5581 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5641 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5701 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5761 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5821 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5881 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5941 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6001 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6061 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6121 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6181 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6241 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6301 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6361 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6421 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6481 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6541 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6601 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6661 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6721 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6781 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6841 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6901 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6961 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7021 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7081 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7141 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7201 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7741 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     7861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8041 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8101 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8161 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8221 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8281 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8341 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8401 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8461 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8521 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8581 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8641 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8701 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8761 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8821 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8881 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8941 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9001 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9061 cacctgacgt c
//
