LOCUS       V022539                 6524 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022539
VERSION     V022539
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..2014
                     /label="PTGER2(NM_000956)"
                     /note="PTGER2(NM_000956)"
                     /gene="PTGER2"
     CDS             2021..2086
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2124..2348
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2394..2822
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2836..3165
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3232..4026
                     /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    4200..4321
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4370..4386)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4394..4410
                     /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(4418..4448)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4463..4484
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4772..5357)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5528..6388)
                     /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(6389..6493)
                     /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 atgggcaatg cctccaatga
      961 ctcccagtct gaggactgcg agacgcgaca gtggcttccc ccaggcgaaa gcccagccat
     1021 cagctccgtc atgttctcgg ccggggtgct ggggaacctc atagcactgg cgctgctggc
     1081 gcgccgctgg cggggggacg tggggtgcag cgccggccgc aggagctccc tctccttgtt
     1141 ccacgtgctg gtgaccgagc tggtgttcac cgacctgctc gggacctgcc tcatcagccc
     1201 agtggtactg gcttcgtacg cgcggaacca gaccctggtg gcactggcgc ccgagagccg
     1261 cgcgtgcacc tacttcgctt tcgccatgac cttcttcagc ctggccacga tgctcatgct
     1321 cttcgccatg gccctggagc gctacctctc gatcgggcac ccctacttct accagcgccg
     1381 cgtctcgcgc tccgggggcc tggccgtgct gcctgtcatc tatgcagtct ccctgctctt
     1441 ctgctcgctg ccgctgctgg actatgggca gtacgtccag tactgccccg ggacctggtg
     1501 cttcatccgg cacgggcgga ccgcttacct gcagctgtac gccaccctgc tgctgcttct
     1561 cattgtctcg gtgctcgcct gcaacttcag tgtcattctc aacctcatcc gcatgcaccg
     1621 ccgaagccgg agaagccgct gcggaccttc cctgggcagt ggccggggcg gccccggggc
     1681 ccgcaggaga ggggaaaggg tgtccatggc ggaggagacg gaccacctca ttctcctggc
     1741 tatcatgacc atcaccttcg ccgtctgctc cttgcctttc acgatttttg catatatgaa
     1801 tgaaacctct tcccgaaagg aaaaatggga cctccaagct cttaggtttt tatcaattaa
     1861 ttcaataatt gacccttggg tctttgccat ccttaggcct cctgttctga gactaatgcg
     1921 ttcagtcctc tgttgtcgga tttcattaag aacacaagat gcaacacaaa cttcctgttc
     1981 tacacagtca gatgccagta aacaggctga ccttctcgag gactacaaag accatgacgg
     2041 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     2101 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2161 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2221 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2281 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2341 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2401 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2461 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2521 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2581 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2641 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2701 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2761 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2821 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3001 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3061 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3121 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3181 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3241 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3301 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3361 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3421 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3481 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3541 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3601 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3661 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3721 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3781 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3841 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3901 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3961 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4021 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4081 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4141 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4201 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4261 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4321 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4381 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4441 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4501 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4561 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4621 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4681 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4741 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4801 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4861 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4921 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4981 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5041 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5101 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5161 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5221 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5281 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6481 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
