LOCUS       V027348                 8804 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027348
VERSION     V027348
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             1660..4176
                     /label="VPS16(NM_022575)"
                     /note="VPS16(NM_022575)"
                     /gene="VPS16"
     CDS             4183..4209
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4213..4239
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4243..4269
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4371..4595
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4641..5069
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5083..5412
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5479..6273
                     /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    6447..6568
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6617..6633)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6641..6657
                     /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(6665..6695)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6710..6731
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7019..7604)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7775..8635)
                     /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(8636..8740)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tggactgcta cacggcgaac
     1681 tggaacccac tcggggactc tgccttttac cggaaatatg agctgtacag catggactgg
     1741 gacctgaagg aggaactcag ggattgcctg gtggctgctg caccctatgg gggccccatt
     1801 gcactgctga ggaacccctg gcggaaggag aaagctgcta gtgtgaggcc agtgctcgat
     1861 atatactctg cttccggcat gcctctggcc agcctgctgt ggaagagtgg acccgtggtg
     1921 tccctgggct ggtcagctga ggaggagctg ctctgtgtgc aggaagatgg tgctgtactg
     1981 gtttatgggc ttcatggtga cttccggaga cacttcagca tgggcaatga agtgctccag
     2041 aaccgggttc tggatgcccg gatctttcac actgagtttg gttccggagt ggccatcctc
     2101 acaggggccc accgcttcac cctcagtgcc aatgtgggtg acctcaaact ccgccggatg
     2161 ccagaggtgc caggtctgca aagtgcaccc tcctgctgga ctgtgctgtg ccaggaccga
     2221 gtggcacaca ttcttctggc tgtggggcct gacctttacc tcttggacca tgcagcctgc
     2281 tccgcagtga cgccccctgg cctggcccca ggagtaagca gcttcctaca gatggctgtc
     2341 tccttcacct accgacacct ggcactcttc acagacacag gctacatctg gatggggaca
     2401 gcatcactca aggagaagct atgtgagttc aactgcaaca tccgggcacc tccaaagcag
     2461 atggtctggt gcagccgtcc tcgtagcaag gagagggccg tggtggtggc ctgggaaagg
     2521 cggctgatgg tggtgggcga tgcacccgag agcatccagt ttgtgctgga tgaggactcc
     2581 tacctggtgc ctgagctcga tggggtccgc atcttctccc gcagcaccca cgagttcctg
     2641 catgaggttc cagcggccag cgaggaaatc ttcaaaattg cctcaatggc ccccggggcg
     2701 ctgctcctgg aggctcagaa ggagtatgag aaagagagcc agaaggcgga cgagtacctg
     2761 cgggagatcc aggagctggg ccagctgacc caggccgtgc agcagtgcat tgaggctgca
     2821 ggacatgagc accagccaga catgcagaag agtctgctca gggccgcctc cttcggaaag
     2881 tgtttcctgg acagatttcc acccgacagc ttcgtgcaca tgtgtcagga cctgcgtgtg
     2941 ctcaatgctg ttcgggacta tcacatcggg atcccgctca cctatagcca atataagcag
     3001 ctcaccatcc aggtgctgct ggacaggctc gtgttgcgga gactttaccc cctggccatc
     3061 cagatatgcg agtacttgcg ccttcctgaa gtacagggcg tcagcaggat cctggcccac
     3121 tgggcctgct acaaggtgca acagaaggat gtctcagatg aggatgtggc tcgagccatt
     3181 aaccagaagc tgggggacac gcctggtgtc tcttactccg acattgctgc acgagcctat
     3241 ggttgtggcc gcacggagct ggccatcaag ctgctggagt atgagccacg ctcaggggag
     3301 caggtacccc ttctcctaaa gatgaagagg agcaaactgg cactaagcaa ggccatcgag
     3361 agcggggaca ctgacctggt gttcacggtg ttgctgcacc tgaagaacga gctgaaccga
     3421 ggagattttt tcatgaccct tcggaatcag cccatggccc tcagtttgta ccgacagttc
     3481 tgtaagcatc aggagctaga gacgctgaag gacctttaca atcaggatga caatcaccag
     3541 gaattgggca gcttccacat ccgagccagc tatgctgcag aagagcgtat tgaggggcga
     3601 gtagcagctc tgcagacagc cgccgatgcc ttctacaagg ccaagaatga gtttgcagcc
     3661 aaggctacag aggatcaaat gcggctccta cggctgcagc ggcgcctaga agacgagctg
     3721 gggggccagt tcctagacct gtctctacat gacacagtta ccaccctcat tcttggcggt
     3781 cacaacaagc gtgcagagca gctggcacgt gacttccgca tccctgacaa gaggctctgg
     3841 tggctgaagc tgactgccct ggcagatttg gaagattggg aagagctaga gaagttttcc
     3901 aagagcaaga aatcacccat tggctacctg ccttttgtgg agatctgcat gaaacaacat
     3961 aacaaatacg aagccaagaa gtatgcttcc cgcgtgggtc ccgagcagaa ggtcaaggct
     4021 ttgcttcttg ttggcgatgt ggctcaggct gcagatgtgg ccatcgaaca ccggaatgag
     4081 gctgagctga gcctcgtatt gtcccactgc acgggagcca cagatggggc cacagctgac
     4141 aagattcaac gggccagggc acaagcccag aagaagggta cctacccata cgacgtacct
     4201 gattatgcaa cttatcctta tgatgtacct gattatgcta gctatcctta tgatgtccca
     4261 gactacgcat aggatccact agtccagtgt ggtggaattc tgcagatatc cagcacagtg
     4321 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     4381 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4441 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4501 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4561 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4621 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4681 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4741 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4801 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4861 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4921 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4981 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5041 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5101 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5161 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5221 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5281 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5341 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5401 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     5461 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5521 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5581 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5641 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5701 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5761 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5821 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5881 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5941 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6001 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6061 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6121 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6181 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6241 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6301 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6361 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6421 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6541 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6601 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6661 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6721 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6781 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6841 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6901 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6961 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7021 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7081 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7141 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7201 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7261 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7321 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7381 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7441 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7501 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     7561 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7621 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7681 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7741 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7801 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7861 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7921 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7981 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8041 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8101 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8161 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8221 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8281 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8341 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8401 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8461 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8521 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8581 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8641 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8701 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8761 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggt
//
