LOCUS       V025960                 7742 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025960
VERSION     V025960
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..3232
                     /label="PIGR(NM_002644)"
                     /note="PIGR(NM_002644)"
                     /gene="PIGR"
     CDS             3239..3304
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3342..3566
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3612..4040
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4054..4383
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4450..5244
                     /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    5418..5539
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5588..5604)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5612..5628
                     /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(5636..5666)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5681..5702
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5990..6575)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6746..7606)
                     /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(7607..7711)
                     /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 atgctgctct tcgtgctcac
      961 ctgcctgctg gcggtcttcc cagccatctc cacgaagagt cccatatttg gtcccgagga
     1021 ggtgaatagt gtggaaggta actcagtgtc catcacgtgc tactacccac ccacctctgt
     1081 caaccggcac acccggaagt actggtgccg gcagggagct agaggtggct gcataaccct
     1141 catctcctcg gagggctacg tctccagcaa atatgcaggc agggctaacc tcaccaactt
     1201 cccggagaac ggcacatttg tggtgaacat tgcccagctg agccaggatg actccgggcg
     1261 ctacaagtgt ggcctgggca tcaatagccg aggcctgtcc tttgatgtca gcctggaggt
     1321 cagccagggt cctgggctcc taaatgacac taaagtctac acagtggacc tgggcagaac
     1381 ggtgaccatc aactgccctt tcaagactga gaatgctcaa aagaggaagt ccttgtacaa
     1441 gcagataggc ctgtaccctg tgctggtcat cgactccagt ggttatgtaa atcccaacta
     1501 tacaggaaga atacgccttg atattcaggg tactggccag ttactgttca gcgttgtcat
     1561 caaccaactc aggctcagcg atgctgggca gtatctctgc caggctgggg atgattccaa
     1621 tagtaataag aagaatgctg acctccaagt gctaaagccc gagcccgagc tggtttatga
     1681 agacctgagg ggctcagtga ccttccactg tgccctgggc cctgaggtgg caaacgtggc
     1741 caaatttctg tgccgacaga gcagtgggga aaactgtgac gtggtcgtca acaccctggg
     1801 gaagagggcc ccagcctttg agggcaggat cctgctcaac ccccaggaca aggatggctc
     1861 attcagtgtg gtgatcacag gcctgaggaa ggaggatgca gggcgctacc tgtgtggagc
     1921 ccattcggat ggtcagctgc aggaaggctc gcctatccag gcctggcaac tcttcgtcaa
     1981 tgaggagtcc acgattcccc gcagccccac tgtggtgaag ggggtggcag gaggctctgt
     2041 ggccgtgctc tgcccctaca accgtaagga aagcaaaagc atcaagtact ggtgtctctg
     2101 ggaaggggcc cagaatggcc gctgccccct gctggtggac agcgaggggt gggttaaggc
     2161 ccagtacgag ggccgcctct ccctgctgga ggagccaggc aacggcacct tcactgtcat
     2221 cctcaaccag ctcaccagcc gggacgccgg cttctactgg tgtctgacca acggcgatac
     2281 tctctggagg accaccgtgg agatcaagat tatcgaagga gaaccaaacc tcaaggtacc
     2341 agggaatgtc acggctgtgc tgggagagac tctcaaggtc ccctgtcact ttccatgcaa
     2401 attctcctcg tacgagaaat actggtgcaa gtggaataac acgggctgcc aggccctgcc
     2461 cagccaagac gaaggcccca gcaaggcctt cgtgaactgt gacgagaaca gccggcttgt
     2521 ctccctgacc ctgaacctgg tgaccagggc tgatgagggc tggtactggt gtggagtgaa
     2581 gcagggccac ttctatggag agactgcagc cgtctatgtg gcagttgaag agaggaaggc
     2641 agcggggtcc cgcgatgtca gcctagcgaa ggcagacgct gctcctgatg agaaggtgct
     2701 agactctggt tttcgggaga ttgagaacaa agccattcag gatcccaggc tttttgcaga
     2761 ggaaaaggcg gtggcagata caagagatca agccgatggg agcagagcat ctgtggattc
     2821 cggcagctct gaggaacaag gtggaagctc cagagcgctg gtctccaccc tggtgcccct
     2881 gggcctggtg ctggcagtgg gagccgtggc tgtgggggtg gccagagccc ggcacaggaa
     2941 gaacgtcgac cgagtttcaa tcagaagcta caggacagac attagcatgt cagacttcga
     3001 gaactccagg gaatttggag ccaatgacaa catgggagcc tcttcgatca ctcaggagac
     3061 atccctcgga ggaaaagaag agtttgttgc caccactgag agcaccacag agaccaaaga
     3121 acccaagaag gcaaaaaggt catccaagga ggaagccgag atggcctaca aagacttcct
     3181 gctccagtcc agcaccgtgg ccgccgaggc ccaggacggc ccccaggaag ccctcgagga
     3241 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3301 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3361 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3421 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3481 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3541 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3601 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3961 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4021 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4201 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4261 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4321 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4381 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4441 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4501 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4561 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4621 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4681 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4741 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4801 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4861 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4921 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4981 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5041 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5101 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5161 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5221 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5281 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5341 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5401 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5461 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5521 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5581 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5641 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5701 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6181 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6421 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7741 tc
//
