LOCUS       V022583                 6914 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022583
VERSION     V022583
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..2404
                     /label="PTGER4(NM_000958)"
                     /note="PTGER4(NM_000958)"
                     /gene="PTGER4"
     CDS             2411..2476
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2514..2738
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2784..3212
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3226..3555
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3622..4416
                     /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    4590..4711
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4760..4776)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4784..4800
                     /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(4808..4838)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4853..4874
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5162..5747)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5918..6778)
                     /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(6779..6883)
                     /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 atgtccactc ccggggtcaa
      961 ttcgtccgcc tccttgagcc ccgaccggct gaacagccca gtgaccatcc cggcggtgat
     1021 gttcatcttc ggggtggtgg gcaacctggt ggccatcgtg gtgctgtgca agtcgcgcaa
     1081 ggagcagaag gagacgacct tctacacgct ggtatgtggg ctggctgtca ccgacctgtt
     1141 gggcactttg ttggtgagcc cggtgaccat cgccacgtac atgaagggcc aatggcccgg
     1201 gggccagccg ctgtgcgagt acagcacctt cattctgctc ttcttcagcc tgtccggcct
     1261 cagcatcatc tgcgccatga gtgtcgagcg ctacctggcc atcaaccatg cctatttcta
     1321 cagccactac gtggacaagc gattggcggg cctcacgctc tttgcagtct atgcgtccaa
     1381 cgtgctcttt tgcgcgctgc ccaacatggg tctcggtagc tcgcggctgc agtacccaga
     1441 cacctggtgc ttcatcgact ggaccaccaa cgtgacggcg cacgccgcct actcctacat
     1501 gtacgcgggc ttcagctcct tcctcattct cgccaccgtc ctctgcaacg tgcttgtgtg
     1561 cggcgcgctg ctccgcatgc accgccagtt catgcgccgc acctcgctgg gcaccgagca
     1621 gcaccacgcg gccgcggccg cctcggttgc ctcccggggc caccccgctg cctccccagc
     1681 cttgccgcgc ctcagcgact ttcggcgccg ccggagcttc cgccgcatcg cgggcgccga
     1741 gatccagatg gtcatcttac tcattgccac ctccctggtg gtgctcatct gctccatccc
     1801 gctcgtggtg cgagtattcg tcaaccagtt atatcagcca agtttggagc gagaagtcag
     1861 taaaaatcca gatttgcagg ccatccgaat tgcttctgtg aaccccatcc tagacccctg
     1921 gatatatatc ctcctgagaa agacagtgct cagtaaagca atagagaaga tcaaatgcct
     1981 cttctgccgc attggcgggt cccgcaggga gcgctccgga cagcactgct cagacagtca
     2041 aaggacatct tctgccatgt caggccactc tcgctccttc atctcccggg agctgaagga
     2101 gatcagcagt acatctcaga ccctcctgcc agacctctca ctgccagacc tcagtgaaaa
     2161 tggccttgga ggcaggaatt tgcttccagg tgtgcctggc atgggcctgg cccaggaaga
     2221 caccacctca ctgaggactt tgcgaatatc agagacctca gactcttcac agggtcagga
     2281 ctcagagagt gtcttactgg tggatgaggc tggtgggagc ggcagggctg ggcctgcccc
     2341 taaggggagc tccctgcaag tcacatttcc cagtgaaaca ctgaacttat cagaaaaatg
     2401 tatactcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     2461 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     2521 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2581 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2641 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2701 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     2761 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2821 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2881 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2941 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3001 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3061 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3121 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3181 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3301 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3361 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3421 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3481 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3541 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3601 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3661 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3721 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3781 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3841 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3901 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3961 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4021 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4081 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4141 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4201 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4261 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4321 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4381 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4441 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4501 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4561 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4741 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4801 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4861 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4921 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5221 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5281 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5341 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5401 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5461 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5521 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5581 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5641 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     5701 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5761 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5821 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5881 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5941 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6001 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6061 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6121 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6181 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6241 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6301 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6361 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6421 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6481 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6541 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6601 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6661 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6721 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6781 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6841 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6901 tgccacctga cgtc
//
