LOCUS       V037078                 7891 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037078
VERSION     V037078
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..3381
                     /label="Mcoln1(NM_053177)"
                     /note="Mcoln1(NM_053177)"
                     /gene="Mcoln1"
     polyA_signal    3458..3682
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3728..4156
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4170..4499
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4566..5360
                     /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    5534..5655
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5704..5720)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5728..5744
                     /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(5752..5782)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5797..5818
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6106..6691)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6862..7722)
                     /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(7723..7827)
                     /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 catggccacc ccggcgggcc ggcgcgcctc agagactgag
     1681 cggctgctga cccccaatcc tgggtatggg acccaggtgg ggacctcacc agccccaaca
     1741 acccccacag aagaggaaga cctccgccgc cgcctcaagt acttctttat gagcccatgt
     1801 gacaagttcc gggccaaagg ccgcaagccc tgcaagctga tgctgcaggt ggtcaagatc
     1861 ttggtggtca ctgtgcagct cattctcttt gggctcagca accagctggt ggtgacattc
     1921 cgggaagaga acaccattgc cttccgacat ctcttcctgc tgggttactc tgatgggtct
     1981 gatgacacct ttgcagccta cacacaggag cagctctacc aagccatctt ctatgctgtg
     2041 gaccagtacc tgatactacc tgagatatcc ctgggccggt atgcctatgt ccgtggtggg
     2101 ggtgggcctt gggccaatgg atcagctttg gctctctgcc agcggtacta ccaccgtggc
     2161 catgtggacc cagccaatga tacctttgac attgatccaa gggtagtcac tgactgtatc
     2221 caggtggatc ctcctgacag accccctgac atccccagtg aggacttgga cttcttggat
     2281 ggcagcgcca gttacaagaa cctcacactg aaattccaca agctgatcaa cgtcaccatc
     2341 cacttccagc tgaagacaat taacctgcag agcctcatca acaatgagat ccctgattgt
     2401 tacaccttca gtatcctgat cacatttgac aataaagcgc acagtgggcg aatccccatc
     2461 cgcctggaga ccaagaccca catccaggag tgcaaacacc ccagtgtctc cagacatgga
     2521 gacaacagct tccggcttct gtttgatgtg gtggttatcc tcacctgctc cctgtccttc
     2581 ctgctgtgcg cccgctcact gctccgtggc ttcctgctgc agaacgagtt tgttgtattc
     2641 atgtggcggc ggcggggtcg ggaaatcagc ctctgggaac ggctggagtt tgtcaatggc
     2701 tggtacatcc tgctggtcac cagtgacgtg ctcaccatct cggggactgt catgaagatt
     2761 ggcattgagg caaagaacct agccagctat gatgtctgca gtattctctt gggtacctcc
     2821 actctgctag tctgggtcgg tgtcattcgc tacctgacat ttttccacaa gtacaacatc
     2881 ttgattgcca cgttgcgagt ggcactgccc agtgtcatgc gtttctgctg ctgtgtggct
     2941 gtcatctacc tgggctattg cttctgtggc tggatcgttc tagggcccta ccatgtgaag
     3001 ttccgctcgc tgtccatggt ttctgagtgt ctgttctcac tcatcaacgg agacgacatg
     3061 tttgtgacgt tcgcggccat gcaggcccag cagggtcaca gcagcctggt gtggctcttc
     3121 tcccagctgt acctctactc cttcatcagc ctcttcatct acatggtgct gagcctcttc
     3181 attgcactca tcaccggcgc ctatgacacc atcaagcacc caggaggtac tggcacagag
     3241 aagagtgagc tccaggccta catcgagcag tgccaggata gccccacatc tggcaagttc
     3301 cgtcgtggca gtggctcagc ttgtagcctt ttctgctgct gtggaaggga ctccccggag
     3361 gaccattcgc tgctggtgaa ctgaggatcc tgatatccag cacagtggcg gccgctcgag
     3421 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3481 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3541 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3601 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3661 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3721 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3781 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3841 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3901 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3961 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4021 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4081 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4141 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4201 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4261 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4321 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4381 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4441 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4501 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4561 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4621 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4681 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4741 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4801 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4861 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4921 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4981 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5041 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5101 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5161 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5221 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5281 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5341 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5401 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5461 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5521 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5581 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5641 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5701 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5761 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5821 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5881 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5941 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6001 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6061 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6121 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6181 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6241 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6301 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6361 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6421 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6481 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6541 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6601 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     6661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6721 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6781 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6841 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6901 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6961 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7021 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7081 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7141 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7201 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7261 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7321 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7381 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7441 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7501 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7561 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7621 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7681 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7741 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7801 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7861 cggatcggga gatctcccga tcccctatgg t
//
