LOCUS       V031965                 7984 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031965
VERSION     V031965
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             890..2695
                     /label="GLS2(NM_013267)"
                     /note="GLS2(NM_013267)"
                     /gene="GLS2"
     CDS             2702..2755
                     /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             2756..3475
                     /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    3551..3775
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3821..4249
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4263..4592
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4659..5453
                     /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    5627..5748
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5797..5813)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5821..5837
                     /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(5845..5875)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5890..5911
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6199..6784)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6955..7815)
                     /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(7816..7920)
                     /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 cttgccacca tgcgctccat
      901 gaaggctctg cagaaggccc tgagccgggc tggcagtcac tgcgggcgag gaggctgggg
      961 tcacccgagc cggagccccc tccttggcgg gggcgtccgg caccacctca gtgaggccgc
     1021 ggcgcagggc agagagacgc cacacagcca ccagccgcag caccaggatc atgattcatc
     1081 agaaagtggc atgctgtccc gcctgggtga tttgctcttt tacactattg ctgaaggaca
     1141 ggaacgaatc cctatccaca agttcaccac tgcactaaag gccactggac tgcagacatc
     1201 agatcctcgg ctccgagact gcatgagcga gatgcaccgc gtggtccaag agtccagtag
     1261 tggtggcctc ttggaccgag atctcttccg aaagtgtgtg agcagcaaca ttgtgctcct
     1321 gacccaggca ttccgaaaga agtttgtcat tcctgatttt gaggagttca cgggccatgt
     1381 ggatcgcatc tttgaggatg tcaaagagct cactggaggc aaagtggcag cctacatccc
     1441 tcagctggcc aagtcaaacc cagacctgtg gggtgtctcc ctgtgcactg tggatggtca
     1501 acggcactct gtgggccaca caaagatccc cttctgcctg cagtcctgtg tgaagcccct
     1561 cacctatgcc atctccataa gcaccctagg cactgactac gtgcacaagt ttgtgggcaa
     1621 agagccaagt ggcctgcgct acaacaagct ctccctcaat gaggaaggaa tcccccataa
     1681 ccccatggtc aatgctggtg ccattgttgt cagctccctg atcaagatgg actgtaacaa
     1741 agcagagaag tttgattttg tgttgcagta tctcaacaaa atggctggga atgaatacat
     1801 gggtttcagc aatgccacat tccagtcaga gaaggaaaca ggggatcgga attatgccat
     1861 cggctattat ctcaaggaaa agaagtgctt tcctaagggg gtggacatga tggctgccct
     1921 tgatctctac ttccagctgt gttctgtgga ggtcacttgt gaatcaggca gtgtcatggc
     1981 agccaccctc gccaacggtg ggatctgccc catcacaggc gagagtgtgc tgagtgctga
     2041 agcagtgcgc aacaccctca gcctcatgca ttcctgcggc atgtatgact tctctggcca
     2101 gtttgccttc cacgtgggcc tgccagccaa gtcagctgta tcaggagcca tcctcctggt
     2161 ggtacccaat gtcatgggaa tgatgtgcct gtcaccccca ttggacaagc tggggaacag
     2221 ccataggggg accagcttct gccagaagtt ggtgtctctc ttcaatttcc acaactatga
     2281 caacctgagg cactgtgctc ggaagttaga cccacggcgt gaaggggcag aaattcggaa
     2341 caagactgtg gtcaacctgt tatttgctgc ctatagtggc gatgtctcag ctcttcgaag
     2401 gtttgccttg tcagccatgg atatggaaca gaaagactat gactcgcgca cagctctgca
     2461 tgttgctgca gctgaaggac acatcgaagt tgttaaattc ctgatcgagg cttgcaaagt
     2521 gaatcctttt gccaaggaca ggtggggcaa cattcccctg gatgatgctg tgcagttcaa
     2581 ccatctggag gtggtcaaac tgcttcaaga ttaccaggac tcctacacac tctctgaaac
     2641 tcaggctgag gcagcagctg aggccctgtc caaagagaac ttagaaagca tggtaggatc
     2701 cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatggt
     2761 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2821 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2881 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2941 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     3001 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3061 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3121 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3181 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3241 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3301 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3361 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3421 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtagaattc
     3481 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     3541 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3601 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3661 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3721 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3781 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3841 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3901 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3961 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4021 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4081 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4141 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4201 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4261 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4381 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4441 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4501 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4561 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4621 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4681 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4741 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4801 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4861 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4921 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4981 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5041 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5101 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5161 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5221 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5281 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5341 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5401 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5461 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5521 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5581 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5641 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5701 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5761 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5821 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5881 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5941 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6001 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6061 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6601 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6721 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7921 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7981 tggt
//
