LOCUS       V038553                 7953 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038553
VERSION     V038553
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             955..2733
                     /label="Miga2(NM_175392)"
                     /note="Miga2(NM_175392)"
                     /gene="Miga2"
     CDS             2749..2802
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             2803..3522
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    3548..3772
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3818..4246
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4260..4589
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4656..5450
                     /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    5626..5747
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5796..5812)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5820..5836
                     /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(5844..5874)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5889..5910
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6198..6786)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6957..7817)
                     /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(7818..7922)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc caccatggca
      961 ttccggagga ctgagggtat gtccatgatc caggctctgg ccatgactgt ggctgagatt
     1021 ccagtgttcc tgtacacaac ttttggtcag tccgcgttct cccagctgcg gttgacaccg
     1081 ggcctgagga aggttctttt tgccacagcc cttgggactg tggccttggc cctggctgcc
     1141 caccagctga aaaggcgccg gcggaagaag aagcaggtgg gccctgagat gggaggtgag
     1201 cagctgggca cggtgcccat gcccatcctc atggcccgca aggtcccttc ggtgaagaaa
     1261 ggctgctcca gcaggagggt tcagagcccc agcagcaaga gcaacgacac tcttagtggc
     1321 atctcctcca tcgagcccag caagcactca ggctcctccc acagcctggc ctcgatggta
     1381 gtggtcaatt catccagccc tacagctgcg tgctcaggtt catgggaagc ccgagggatg
     1441 gaggagtctg tgcccaccac tgatggcagt gccgagagcc tctatgtgca aggcatggag
     1501 ctgtttgagg aagccctgca gaaatgggag caggcactga gtgtgggcca gagaggggat
     1561 ggaggcagca cccccacacc aggcgacagc ctccagaacc cagacaccgc atcagaagca
     1621 ctgtcagagc cagagtccca acgaagggag tttgctgaga agctggagtc cctgctgcac
     1681 cgggcctacc acctgcagga agagtttggc tctaccttcc cctctgacag catgctgttg
     1741 gatcttgaga ggaccctaat gctgccactg actgagggct cactgcgact tcgagctgac
     1801 gatgaggaca gcctgacctc tgaagactcc tttttctcag ccactgagat ctttgagtcc
     1861 ctgcagatcg gcgagtaccc gcttcctctc tccaggcctg ctgctgccta cgaggaggct
     1921 ctgcagctgg tgaaggaagg aagagtccct tgccgaacac tcaggacaga gctgctgggc
     1981 tgctacagtg accaggactt cctagccaag ctgcattgtg tacggcaggc ctttgagggg
     2041 cttctagaag aaagaagcaa ccagatcttc tttggggagg tcggccggca gatggtgacg
     2101 ggcctgatga ccaaggctga gaagagtccc aaaggcttcc tggagagcta tgaggagatg
     2161 ctgagctacg ccctgcggcc tgagacctgg gctaccaccc ggctggagct ggagggcaga
     2221 ggggtggcct gcatgagctt cttcgacatc gtgctggact tcatcctcat ggatgccttt
     2281 gaggacctag agaaccctcc gtcttcggtg cttgctgtcc tgaggaaccg ctggctgtct
     2341 gacagcttca aagaaacagc cctagccacc gcgtgctggt ctgtcctgaa agccaagagg
     2401 aggctcctga tggtgcctga tggcttcatc tcccatttct actccgtatc ggagcatgtt
     2461 agccctgtgt tggcctttgg cttcctcgga cccaagcccc agctctcgga agtctgtgct
     2521 ttcttcaagc accagattgt acaatacctg agggacatgt tcgacctgga caacgtgcgc
     2581 tacacatctg tgccagccct agcagaagac atcctacagc tgtcccggcg gcgcagcgag
     2641 attctattgg gctacctggg ggcacctgtg gctagtagca tcgggctgaa tgggccactg
     2701 cctcgagaaa atgggcccct ggaagagctg cagtctagag ggcccttcga gggcagagga
     2761 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatggtgag caagggcgag
     2821 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2881 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2941 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3001 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3061 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3121 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3181 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3241 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3301 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3361 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3421 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3481 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtttaaac ccgctgatca
     3541 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3601 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3661 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3721 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3781 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3841 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3901 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3961 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4021 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4081 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4141 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4201 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4261 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4381 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4441 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4501 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4561 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4621 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4681 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4741 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4801 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4861 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4921 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4981 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5041 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5101 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5161 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5221 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5281 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5341 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5401 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5461 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5521 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5581 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5641 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5701 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5761 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5821 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5881 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5941 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6001 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6061 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6121 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6601 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6961 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7021 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7081 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7141 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7201 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7261 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7321 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7381 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7441 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7501 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7561 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7621 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7681 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7741 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7801 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7861 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7921 cgcacatttc cccgaaaagt gccacctgac gtc
//
