LOCUS       V025670                 8581 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025670
VERSION     V025670
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             874..1590
                     /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"
     CDS             1642..4071
                     /label="LRRC8A(NM_001127244)"
                     /note="LRRC8A(NM_001127244)"
                     /gene="LRRC8A"
     polyA_signal    4148..4372
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4418..4846
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4860..5189
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5256..6050
                     /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    6224..6345
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6394..6410)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6418..6434
                     /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(6442..6472)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6487..6508
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6796..7381)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7552..8412)
                     /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(8413..8517)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgattccg gtgacagagc tccgctactt tgcggacacg
     1681 cagccagcat accggatcct gaagccgtgg tgggatgtgt tcacagacta catctctatc
     1741 gtcatgctga tgattgccgt cttcgggggg acgctgcagg tcacccaaga caagatgatc
     1801 tgcctgcctt gtaagtgggt caccaaggac tcctgcaatg attcgttccg gggctgggca
     1861 gcccctggcc cggagcccac ctaccccaac tccaccattc tgccgacccc tgacacgggc
     1921 cccacaggca tcaagtatga cctggaccgg caccagtaca actacgtgga cgctgtgtgc
     1981 tatgagaacc gactgcactg gtttgccaag tacttcccct acctggtgct tctgcacacg
     2041 ctcatcttcc tggcctgcag caacttctgg ttcaaattcc cgcgcaccag ctcgaagctg
     2101 gagcactttg tgtctatcct gctgaagtgc ttcgactcgc cctggaccac gagggccctg
     2161 tcggagacag tggtggagga gagcgacccc aagccggcct tcagcaagat gaatgggtcc
     2221 atggacaaaa agtcatcgac cgtcagtgag gacgtggagg ccaccgtgcc catgctgcag
     2281 cggaccaagt cacggatcga gcagggtatc gtggaccgct cagagacggg cgtgctggac
     2341 aagaaggagg gggagcaagc caaggcgctg tttgagaagg tgaagaagtt ccggacccat
     2401 gtggaggagg gggacattgt gtaccgcctc tacatgcggc agaccatcat caaggtgatc
     2461 aagttcatcc tcatcatctg ctacaccgtc tactacgtgc acaacatcaa gttcgacgtg
     2521 gactgcaccg tggacattga gagcctgacg ggctaccgca cctaccgctg tgcccacccc
     2581 ctggccacac tcttcaagat cctggcgtcc ttctacatca gcctagtcat cttctacggc
     2641 ctcatctgca tgtatacact gtggtggatg ctacggcgct ccctcaagaa gtactcgttt
     2701 gagtcgatcc gtgaggagag cagctacagc gacatccccg acgtcaagaa cgacttcgcc
     2761 ttcatgctgc acctcattga ccaatacgac ccgctctact ccaagcgctt cgccgtcttc
     2821 ctgtcggagg tgagtgagaa caagctgcgg cagctgaacc tcaacaacga gtggacgctg
     2881 gacaagctcc ggcagcggct caccaagaac gcgcaggaca agctggagct gcacctgttc
     2941 atgctcagtg gcatccctga cactgtgttt gacctggtgg agctggaggt cctcaagctg
     3001 gagctgatcc ccgacgtgac catcccgccc agcattgccc agctcacggg cctcaaggag
     3061 ctgtggctct accacacagc ggccaagatt gaagcgcccg cgctggcctt cctgcgtgag
     3121 aacctgcggg cgctgcacat caagttcacc gacatcaagg agatcccgct gtggatctat
     3181 agcctgaaga cactggagga gctgcacctg acgggcaacc tgagcgcgga gaacaaccgc
     3241 tacatcgtca tcgacgggct gcgggagctc aaacgcctca aggtgctgcg gctcaagagc
     3301 aacctaagca agctgccaca ggtggtcaca gatgtgggcg tgcacctgca gaagctgtcc
     3361 atcaacaatg agggcaccaa gctcatcgtc ctcaacagcc tcaagaagat ggcgaacctg
     3421 actgagctgg agctgatccg ctgtgacctg gagcgcatcc cccactccat cttcagcctc
     3481 cacaacctgc aggagattga cctcaaggac aacaacctca agaccatcga ggagatcatc
     3541 agcttccagc acctgcaccg cctcacctgc cttaagctgt ggtacaacca catcgcctac
     3601 atccccatcc agatcggcaa cctcaccaac ctggagcgcc tctacctgaa ccgcaacaag
     3661 atcgagaaga tccccaccca gctcttctac tgccgcaagc tgcgctacct ggacctcagc
     3721 cacaacaacc tgaccttcct ccctgccgac atcggcctcc tgcagaacct ccagaaccta
     3781 gccatcacgg ccaaccggat cgagacgctc cctccggagc tcttccagtg ccggaagctg
     3841 cgggccctgc acctgggcaa caacgtgctg cagtcactgc cctccagggt gggcgagctg
     3901 accaacctga cgcagatcga gctgcggggc aaccggctgg agtgcctgcc tgtggagctg
     3961 ggcgagtgcc cactgctcaa gcgcagcggc ttggtggtgg aggaggacct gttcaacaca
     4021 ctgccacccg aggtgaagga gcggctgtgg agggctgaca aggagcaggc ctgaggatcc
     4081 tgatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     4141 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4201 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4261 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4321 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4381 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4441 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4501 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4561 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4621 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4681 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4741 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4801 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4861 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4921 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4981 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5041 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5101 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5161 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5221 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5281 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5341 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5401 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5461 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5521 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5581 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5641 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5701 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5761 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5821 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5881 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5941 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6001 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6061 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6121 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6181 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6241 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6301 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6361 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6421 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6481 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7321 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7381 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7441 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7501 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7561 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7621 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7681 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7741 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7801 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7861 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7921 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7981 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8041 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8101 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8161 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8221 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8281 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8341 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8401 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8461 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8521 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8581 t
//
