LOCUS       V032860                 8259 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032860
VERSION     V032860
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..2939
                     /label="Hgf(NM_001289458)"
                     /note="Hgf(NM_001289458)"
                     /gene="Hgf"
     CDS             2946..3011
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3035..3623
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3689..3737
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3939..4367
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4381..4710
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4777..5571
                     /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    5747..5868
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5917..5933)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5941..5957
                     /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(5965..5995)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6010..6031
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6319..6907)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7078..7938)
                     /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(7939..8043)
                     /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 ccaccatgat gtgggggacc aaacttctgc
      781 cggtcctgtt gctgcagcat gtcctcctgc acctcctcct gcttcatgtc gccatcccct
      841 atgcagaagg acagaagaaa agaagaaata cacttcatga atttaaaaag tcagcaaaaa
      901 ctactcttac caaggaagac ccattactga agattaaaac caaaaaagtg aactctgcag
      961 atgagtgtgc caacaggtgt atcaggaaca ggggctttac gttcacttgc aaggccttcg
     1021 tttttgataa gtcaagaaaa cgatgctact ggtatccttt caatagtatg tcaagtggag
     1081 tgaaaaaagg gtttggccat gaatttgacc tctatgaaaa caaagactat attagaaact
     1141 gcatcattgg taaaggaggc agctataaag ggacggtatc catcactaag agtggcatca
     1201 aatgccagcc ttggaattcc atgatccccc atgaacacag ctttttgcct tcgagctatc
     1261 gcggtaaaga cctacaggaa aactactgtc gaaatcctcg aggggaagaa gggggaccct
     1321 ggtgtttcac aagcaatcca gaggtacgct acgaagtctg tgacattcct cagtgttcag
     1381 aagttgaatg catgacctgc aatggtgaaa gctacagagg tcccatggat cacacagaat
     1441 caggcaagac ttgtcagcgc tgggaccagc agacaccaca ccggcacaag ttcttgccag
     1501 aaagatatcc cgacaagggc tttgatgata attattgccg caatcctgat ggcaagccga
     1561 ggccatggtg ctacactctt gaccctgaca ccccttggga gtattgtgca attaaaacgt
     1621 gcgctcacag tgctgtgaat gagactgatg tccctatgga aacaactgaa tgcattcaag
     1681 gccaaggaga aggttacagg ggaaccagca ataccatttg gaatggaatt ccctgtcagc
     1741 gttgggattc gcagtaccct cacaagcatg atatcactcc cgagaacttc aaatgcaagg
     1801 accttagaga aaattattgc cgcaatccag atggggctga atcaccatgg tgttttacca
     1861 ctgacccaaa catccgagtt ggctactgct ctcaaattcc caagtgtgac gtgtcaagtg
     1921 gacaagattg ttatcgtggc aatgggaaaa attacatggg caacttatcc aaaacaaggt
     1981 ctggacttac atgttccatg tgggacaaga atatggagga tttacaccgt catatcttct
     2041 gggagccaga tgctagcaaa ttgaataaga attactgccg gaatcctgat gatgatgccc
     2101 atggaccttg gtgctacacg gggaatcctc ttattccttg ggattattgc cctatttccc
     2161 gttgtgaagg agatactaca cctacaattg tcaatttgga ccatcctgta atatcctgtg
     2221 ccaaaacaaa acaactgcgg gttgtaaatg gcattccaac acaaacaaca gtagggtgga
     2281 tggttagttt gaaatacaga aataaacata tctgtggagg atcattgata aaggaaagtt
     2341 gggttcttac tgcaagacaa tgttttccag ccagaaacaa agacttgaaa gactatgaag
     2401 cttggcttgg catccacgat gttcatgaga gaggcgagga gaagcgcaag cagatcttaa
     2461 acatttccca gctggtctat ggtcctgaag gctcagactt ggttttactg aagcttgctc
     2521 gacctgcaat cctggataac tttgtcagta caattgattt acctagttat ggttgtacaa
     2581 tccctgaaaa gaccacttgc agtatttacg gctggggcta cactggattg atcaacgcgg
     2641 atggtttatt acgagtagct catctgtata ttatggggaa tgagaaatgc agtcagcacc
     2701 atcaaggcaa ggtgactttg aatgagtctg agttatgtgc tggggctgaa aagattggat
     2761 caggaccatg tgagggagat tatggtggcc cactcatttg tgaacaacac aaaatgagaa
     2821 tggttcttgg tgtcattgtt cctggtcgtg gatgtgccat cccaaatcgt cctggtattt
     2881 ttgttcgagt agcatattat gcaaaatgga tacacaaagt aattttgaca tacaagttgg
     2941 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3001 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     3061 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3121 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3181 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3241 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3301 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3361 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3421 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3481 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3541 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3601 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3661 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3721 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3781 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3841 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3901 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3961 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4021 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4081 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4141 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4201 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4261 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4321 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4381 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4441 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4501 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4561 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4621 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4681 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4741 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4801 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4861 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4921 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4981 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5041 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5101 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5161 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5221 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5281 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5341 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5401 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5461 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5521 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5581 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5641 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5701 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5761 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5821 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5881 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5941 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6001 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6061 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6121 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6481 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6721 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8041 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8101 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8161 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8221 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
