LOCUS       V029202                 7507 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029202
VERSION     V029202
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2943
                     /label="AMER2(NM_152704)"
                     /note="AMER2(NM_152704)"
                     /gene="AMER2"
     CDS             2950..3039
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3074..3298
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3344..3772
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3786..4115
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4182..4976
                     /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    5150..5271
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5320..5336)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5344..5360
                     /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(5368..5398)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5413..5434
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5722..6307)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6478..7338)
                     /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(7339..7443)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggagacga gccggagccg cggcggcggc
      961 ggggctgtca gcgagcgcgg cggagctggc gcgtccgtgg gggtctgcag gaggaaggcg
     1021 gaggccgggg ccgggaccgg gaccctcgcg gcagacatgg acttgcattg tgactgtgcc
     1081 gccgaaacgc cggccgccga gccgccgtcg gggaagatta ataaagctgc cttcaaatta
     1141 ttcaagaaga ggaaatcggg tggcaccatg cccagcattt ttggggtcaa aaacaaaggg
     1201 gacgggaaaa gctcgggtcc gacggggctg gtgaggagca ggacccacga cggacttgcc
     1261 gaggtgctgg tgctggagag cggcaggaag gaggagccgc gcggcggggg cgacagcggc
     1321 gggggcggcg gggggcggcc gaacccgggg ccccccagag ccgcagggcc cggcgggggc
     1381 tccctcgcca gcagctcggt ggccaagtcg cacagcttct tctcgctgct gaagaagaac
     1441 gggcgctcgg aaaacggcaa gggagagcct gtggacgcga gcaaggccgg cggcaaacaa
     1501 aagcgggggc tgcgggggct gttcagcggc atgcgctggc acaggaaaga caagcgggcc
     1561 aaggcggagg ccgcggaggg gcgcgcgccc gggggcggct tgatcctacc cggctcgctc
     1621 accgccagcc tggagtgcgt caaggaggag acgcccagag ccgcgcgcga gccggaggag
     1681 cccagccagg acgccccgcg agacccagca ggtgagcccg cagggggaga ggaggtgccc
     1741 gcccccgccg accgcgcccc agcgcggagc tgccgagagg cagagggcct cgcgcacccc
     1801 ggcgacaccg gcgcccgggg agaggacgcc gcggggcatc ggcgcgccga gccggggccc
     1861 ggggaggtcc gcacggcaga ggacgcttcc aggacggggg ccgttcccgt aaagacggtc
     1921 ccccttgtcg actccgaagg cggcagcggc cgggcgcccg ccgccccaga ccctgcctct
     1981 gtcgatccac cctcagaccc gtcggcagat cgtatttgtt tgatgttttc tgacgtgact
     2041 tcactgaaaa gctttgactc tcttacaggc tgtggagata ttattgcaga ccaagaggaa
     2101 gaggcaggtc ccagctgtga caagcatgtc cccgggccag gcaagccggc tctgtctaaa
     2161 aagaaccccg gcgtggtggc ctaccaagga ggcggggaag agatggccag cccggacgag
     2221 gtggacgaca cctatctaca ggagttctgg gacatgctct cccagaccga ggagcaggga
     2281 cccgagcccc aggagggcgc ggctaaggtg gcagctgcgc tggaaaccaa ggtggtgccc
     2341 gagaccccca aagacaccag gtgtgtggaa gcggccaagg acgcgtcctc ggtcaagcgc
     2401 aggaggctca accggattcc catcgagccc catcctaagg aggagcccaa gcacccggag
     2461 aaggagcagc aggaaggcgt ccccaacagc gacgagggct actgggactc caccacgcca
     2521 ggcccagagg aagacagctc gagcagcggg aagaaggcgg gcatcccccg ggatagctac
     2581 agcggggacg cgctctatga tctctatgct gacccggacg gaagtccagc aacccttcct
     2641 ggagggaagg acaacgagga gacgtcctcc ctgtcccggt taaagcccgt atctccaggc
     2701 accatcacct gtccactgcg aacaccaggc agcttgctga aggactctaa gatccctatt
     2761 agcatcaagc acctgaccaa ccttccatct agccatcccg tggtgcacca gcaaccctcc
     2821 aggagtgaga tgcccagaac aaaaatcccg gtttccaaag tgctggtccg cagagtcagc
     2881 aaccggggct tggctgggac caccatcaga gcaacggcct gccacgacag tgccaaaaag
     2941 ttggatatcg aacaaaaact aatatcagag gaagatttgg aacagaaact aatcagtgaa
     3001 gaagatttag agcaaaagtt aatttccgag gaggacttat gagggcccgt ttaaacccgc
     3061 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3121 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3181 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3241 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3301 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3361 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3421 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3481 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3541 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3601 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3661 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3721 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3781 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3841 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3961 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4021 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4081 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4141 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4201 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4261 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4321 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4381 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4441 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4501 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4561 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4621 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4681 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4741 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4801 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4861 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4921 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4981 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5041 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5101 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5161 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5221 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5281 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5341 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5401 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5461 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5521 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5581 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5641 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5701 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5761 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5821 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5881 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5941 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6001 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6061 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6121 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6181 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6241 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7441 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7501 ctatggt
//
