LOCUS       V040701                 7522 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040701
VERSION     V040701
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             964..3012
                     /label="TGFBI(NM_000358)"
                     /note="TGFBI(NM_000358)"
                     /gene="TGFBI"
     CDS             3019..3084
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3122..3346
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3392..3820
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3834..4163
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4230..5024
                     /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    5198..5319
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5368..5384)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5392..5408
                     /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(5416..5446)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5461..5482
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5770..6355)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6526..7386)
                     /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(7387..7491)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggcgc tcttcgtgcg gctgctggct ctcgccctgg ctctggccct gggccccgcc
     1021 gcgaccctgg cgggtcccgc caagtcgccc taccagctgg tgctgcagca cagcaggctc
     1081 cggggccgcc agcacggccc caacgtgtgt gctgtgcaga aggttattgg cactaatagg
     1141 aagtacttca ccaactgcaa gcagtggtac caaaggaaaa tctgtggcaa atcaacagtc
     1201 atcagctacg agtgctgtcc tggatatgaa aaggtccctg gggagaaggg ctgtccagca
     1261 gccctaccac tctcaaacct ttacgagacc ctgggagtcg ttggatccac caccactcag
     1321 ctgtacacgg accgcacgga gaagctgagg cctgagatgg aggggcccgg cagcttcacc
     1381 atcttcgccc ctagcaacga ggcctgggcc tccttgccag ctgaagtgct ggactccctg
     1441 gtcagcaatg tcaacattga gctgctcaat gccctccgct accatatggt gggcaggcga
     1501 gtcctgactg atgagctgaa acacggcatg accctcacct ctatgtacca gaattccaac
     1561 atccagatcc accactatcc taatgggatt gtaactgtga actgtgcccg gctgctgaaa
     1621 gccgaccacc atgcaaccaa cggggtggtg cacctcatcg ataaggtcat ctccaccatc
     1681 accaacaaca tccagcagat cattgagatc gaggacacct ttgagaccct tcgggctgct
     1741 gtggctgcat cagggctcaa cacgatgctt gaaggtaacg gccagtacac gcttttggcc
     1801 ccgaccaatg aggccttcga gaagatccct agtgagactt tgaaccgtat cctgggcgac
     1861 ccagaagccc tgagagacct gctgaacaac cacatcttga agtcagctat gtgtgctgaa
     1921 gccatcgttg cggggctgtc tgtagagacc ctggagggca cgacactgga ggtgggctgc
     1981 agcggggaca tgctcactat caacgggaag gcgatcatct ccaataaaga catcctagcc
     2041 accaacgggg tgatccacta cattgatgag ctactcatcc cagactcagc caagacacta
     2101 tttgaattgg ctgcagagtc tgatgtgtcc acagccattg accttttcag acaagccggc
     2161 ctcggcaatc atctctctgg aagtgagcgg ttgaccctcc tggctcccct gaattctgta
     2221 ttcaaagatg gaacccctcc aattgatgcc catacaagga atttgcttcg gaaccacata
     2281 attaaagacc agctggcctc taagtatctg taccatggac agaccctgga aactctgggc
     2341 ggcaaaaaac tgagagtttt tgtttatcgt aatagcctct gcattgagaa cagctgcatc
     2401 gcggcccacg acaagagggg gaggtacggg accctgttca cgatggaccg ggtgctgacc
     2461 cccccaatgg ggactgtcat ggatgtcctg aagggagaca atcgctttag catgctggta
     2521 gctgccatcc agtctgcagg actgacggag accctcaacc gggaaggagt ctacacagtc
     2581 tttgctccca caaatgaagc cttccgagcc ctgccaccaa gagaacggag cagactcttg
     2641 ggagatgcca aggaacttgc caacatcctg aaataccaca ttggtgatga aatcctggtt
     2701 agcggaggca tcggggccct ggtgcggcta aagtctctcc aaggtgacaa gctggaagtc
     2761 agcttgaaaa acaatgtggt gagtgtcaac aaggagcctg ttgccgagcc tgacatcatg
     2821 gccacaaatg gcgtggtcca tgtcatcacc aatgttctgc agcctccagc caacagacct
     2881 caggaaagag gggatgaact tgcagactct gcgcttgaga tcttcaaaca agcatcagcg
     2941 ttttccaggg cttcccagag gtctgtgcga ctagcccctg tctatcaaaa gttattagag
     3001 aggatgaagc atctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc
     3061 gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg atcagcctcg
     3121 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3181 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3241 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3301 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3361 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3421 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3481 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3541 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3601 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3661 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3721 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3781 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3841 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3901 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3961 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4021 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4081 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4141 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4201 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4261 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4321 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4381 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4441 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4501 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4561 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4621 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4681 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4741 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4801 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4861 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4921 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4981 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5041 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5101 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5161 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5221 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5281 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5341 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5401 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5461 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5521 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5581 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5641 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5701 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5761 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5821 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5881 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5941 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6001 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6061 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6181 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6301 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7501 ccgaaaagtg ccacctgacg tc
//
