LOCUS       V037799                 8778 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037799
VERSION     V037799
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             927..3584
                     /label="VPS39(NM_001301138)"
                     /note="VPS39(NM_001301138)"
                     /gene="VPS39"
     CDS             3621..4340
                     /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    4378..4602
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4648..5076
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5090..5419
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5486..6280
                     /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    6454..6575
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6624..6640)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6648..6664
                     /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(6672..6702)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6717..6738
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7026..7611)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7782..8642)
                     /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(8643..8747)
                     /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 gctggctagc
      901 gtttaaagcc actgggatcc gccaccatgc acgacgcttt cgagccagtg ccgatcctag
      961 aaaagctgcc tctgcaaatc gactgtctgg ctgcctggga ggaatggctt cttgtgggaa
     1021 ccaaacaagg acatcttctt ctctatagga ttcggaagga cgttgtgcca gcagatgtag
     1081 catcacctga aagcggcagt tgcaacagat ttgaagtgac actagagaaa tccaataaga
     1141 acttctccaa aaagattcag cagatccatg tggtttccca gtttaagatt ctggtcagct
     1201 tgttagaaaa taacatttat gtccatgacc tattgacatt tcaacaaatc actacggttt
     1261 caaaggcaaa gggagcatca ctgtttactt gtgacctcca gcacacagag accggtgagg
     1321 aggtgttacg gatgtgtgtg gcagtaaaaa agaagctgca gctctatttc tggaaggaca
     1381 gggaatttca tgaattgcag ggggacttta gtgtgccaga tgtgcccaag tccatggcgt
     1441 ggtgtgaaaa ttctatctgt gtgggtttca agagagacta ctacctaata agggtggatg
     1501 gaaaggggtc catcaaagag ctctttccaa caggaaaaca gctggagccc ttagttgcac
     1561 ctctggcaga tggaaaagtg gctgtgggcc aggatgatct caccgtggta ctcaatgagg
     1621 aagggatctg cacacagaaa tgtgccctga actggacgga cataccagtg gccatggagc
     1681 accagcctcc ctacatcatt gcagtgttgc ctcgatatgt tgagatccga acatttgaac
     1741 cgaggcttct ggtccaaagc attgaattgc aaaggccccg tttcattacc tcaggaggat
     1801 caaacattat ctatgtggcc agcaatcatt ttgtttggag actcatccct gtccccatgg
     1861 caacccaaat ccaacaactt ctccaggaca agcagtttga attggctctg cagctcgcag
     1921 aaatgaaaga tgattctgac agtgaaaagc agcaacaaat tcatcacatc aagaacttgt
     1981 atgccttcaa cctcttctgc cagaagcgtt ttgatgagtc catgcaggtc tttgctaaac
     2041 ttggcacaga tcccacccat gtgatgggcc tgtaccctga cctgctgccc acagactaca
     2101 gaaagcagtt gcagtatccc aacccattgc ctgtgctctc cggggctgaa ttggagaagg
     2161 ctcacttagc tctgattgac tacctgacac agaaacgaag tcaattggta aagaagctga
     2221 atgactctga tcaccagtca agcacctcac cgctcatgga aggcactccc accatcaaat
     2281 ccaagaagaa gctgctacaa atcatcgaca ccaccctgct caagtgctat ctccatacaa
     2341 atgtggccct ggtggccccc ttgctacgcc tggagaacaa tcactgccac atcgaggaga
     2401 gcgagcacgt gctaaagaag gctcacaagt acagtgagct tatcatcctg tatgagaaga
     2461 aggggctcca cgagaaagct ctgcaggtgc tcgtggacca gtccaagaaa gccaactccc
     2521 ctctgaaagg ccacgagagg acagtgcagt atctgcagca tctgggcaca gaaaacctgc
     2581 atttgatttt ctcctactca gtgtgggtgc tgagagactt cccagaagat ggcctgaaga
     2641 tatttactga agatctcccg gaagtggagt ctctgccacg tgatcgagtc ctcggcttct
     2701 taatagagaa ttttaagggt ctggctattc cttatctgga acacatcatc catgtttggg
     2761 aggagacagg ctctcggttc cacaactgcc tgatccagct atactgtgag aaggtgcaag
     2821 gtctgatgaa ggagtatctc ctgtccttcc ctgcaggcaa aaccccagtc ccagctggag
     2881 aggaagaggg tgagctggga gaataccggc aaaagctcct catgttcttg gagatttcca
     2941 gctactatga tccaggccgg ctcatctgtg attttccctt tgatggcctc ttagaagaac
     3001 gagctctcct gttggggcgc atggggaaac atgaacaagc tcttttcatt tatgtccaca
     3061 tcttgaagga tacaaggatg gctgaggagt actgccacaa acactatgac cgaaacaaag
     3121 atggcaacaa agatgtgtat ctgtccctgc ttcggatgta cctgtcgccc cccagcattc
     3181 actgcctggg gccaatcaag ctggaactac tggagccaaa agccaacctc caggccgctc
     3241 tgcaggtcct cgagctacac cacagcaaac tggacaccac caaggccctc aaccttctgc
     3301 cagcaaacac tcagatcaat gacatacgca tcttcctgga aaaggtcttg gaagaaaatg
     3361 cacaaaagaa acggttcaat caagtgctca agaaccttct ccatgcagaa ttcctgaggg
     3421 tccaggaaga gcggatttta caccagcagg tgaagtgcat catcacagag gagaaggtgt
     3481 gcatggtgtg taagaagaag attgggaaca gtgcatttgc aagatacccc aatggagtgg
     3541 tcgtccatta cttctgttcc aaagaggtaa acccagctga cactctcgag ggcggtggag
     3601 gaagcggcgg aggtggatct atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3661 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3721 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3781 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3841 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3901 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3961 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     4021 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     4081 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     4141 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4201 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     4261 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4321 tggacgagct gtacaagtag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     4381 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4441 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4501 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4561 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4621 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4681 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4741 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4801 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4861 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4921 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4981 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5041 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     5101 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5161 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5221 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5281 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5341 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     5401 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5461 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5521 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5581 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5641 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5701 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5761 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5821 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5881 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5941 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6001 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6061 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6121 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6181 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6241 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6301 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6361 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6421 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     6481 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6541 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     6601 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6661 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6721 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6781 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6841 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6901 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6961 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7021 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7081 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7141 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7201 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7261 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7321 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7381 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7441 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7501 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7561 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7621 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7681 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7741 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7801 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7861 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7921 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7981 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8041 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8101 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8161 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8221 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8281 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8341 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8401 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8461 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8521 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8581 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8641 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8701 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8761 aaagtgccac ctgacgtc
//
