LOCUS       V047604                 8783 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047604
VERSION     V047604
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             884..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1628..4285
                     /label="VPS39(NM_001301138)"
                     /note="VPS39(NM_001301138)"
                     /gene="VPS39"
     CDS             4286..4303
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    4350..4574
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4620..5048
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5062..5391
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5458..6252
                     /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    6426..6547
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6596..6612)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6620..6636
                     /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(6644..6674)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6689..6710
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6998..7583)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7754..8614)
                     /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(8615..8719)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg cacgacgctt tcgagccagt gccgatccta gaaaagctgc ctctgcaaat
     1681 cgactgtctg gctgcctggg aggaatggct tcttgtggga accaaacaag gacatcttct
     1741 tctctatagg attcggaagg acgttgtgcc agcagatgta gcatcacctg aaagcggcag
     1801 ttgcaacaga tttgaagtga cactagagaa atccaataag aacttctcca aaaagattca
     1861 gcagatccat gtggtttccc agtttaagat tctggtcagc ttgttagaaa ataacattta
     1921 tgtccatgac ctattgacat ttcaacaaat cactacggtt tcaaaggcaa agggagcatc
     1981 actgtttact tgtgacctcc agcacacaga gaccggtgag gaggtgttac ggatgtgtgt
     2041 ggcagtaaaa aagaagctgc agctctattt ctggaaggac agggaatttc atgaattgca
     2101 gggggacttt agtgtgccag atgtgcccaa gtccatggcg tggtgtgaaa attctatctg
     2161 tgtgggtttc aagagagact actacctaat aagggtggat ggaaaggggt ccatcaaaga
     2221 gctctttcca acaggaaaac agctggagcc cttagttgca cctctggcag atggaaaagt
     2281 ggctgtgggc caggatgatc tcaccgtggt actcaatgag gaagggatct gcacacagaa
     2341 atgtgccctg aactggacgg acataccagt ggccatggag caccagcctc cctacatcat
     2401 tgcagtgttg cctcgatatg ttgagatccg aacatttgaa ccgaggcttc tggtccaaag
     2461 cattgaattg caaaggcccc gtttcattac ctcaggagga tcaaacatta tctatgtggc
     2521 cagcaatcat tttgtttgga gactcatccc tgtccccatg gcaacccaaa tccaacaact
     2581 tctccaggac aagcagtttg aattggctct gcagctcgca gaaatgaaag atgattctga
     2641 cagtgaaaag cagcaacaaa ttcatcacat caagaacttg tatgccttca acctcttctg
     2701 ccagaagcgt tttgatgagt ccatgcaggt ctttgctaaa cttggcacag atcccaccca
     2761 tgtgatgggc ctgtaccctg acctgctgcc cacagactac agaaagcagt tgcagtatcc
     2821 caacccattg cctgtgctct ccggggctga attggagaag gctcacttag ctctgattga
     2881 ctacctgaca cagaaacgaa gtcaattggt aaagaagctg aatgactctg atcaccagtc
     2941 aagcacctca ccgctcatgg aaggcactcc caccatcaaa tccaagaaga agctgctaca
     3001 aatcatcgac accaccctgc tcaagtgcta tctccataca aatgtggccc tggtggcccc
     3061 cttgctacgc ctggagaaca atcactgcca catcgaggag agcgagcacg tgctaaagaa
     3121 ggctcacaag tacagtgagc ttatcatcct gtatgagaag aaggggctcc acgagaaagc
     3181 tctgcaggtg ctcgtggacc agtccaagaa agccaactcc cctctgaaag gccacgagag
     3241 gacagtgcag tatctgcagc atctgggcac agaaaacctg catttgattt tctcctactc
     3301 agtgtgggtg ctgagagact tcccagaaga tggcctgaag atatttactg aagatctccc
     3361 ggaagtggag tctctgccac gtgatcgagt cctcggcttc ttaatagaga attttaaggg
     3421 tctggctatt ccttatctgg aacacatcat ccatgtttgg gaggagacag gctctcggtt
     3481 ccacaactgc ctgatccagc tatactgtga gaaggtgcaa ggtctgatga aggagtatct
     3541 cctgtccttc cctgcaggca aaaccccagt cccagctgga gaggaagagg gtgagctggg
     3601 agaataccgg caaaagctcc tcatgttctt ggagatttcc agctactatg atccaggccg
     3661 gctcatctgt gattttccct ttgatggcct cttagaagaa cgagctctcc tgttggggcg
     3721 catggggaaa catgaacaag ctcttttcat ttatgtccac atcttgaagg atacaaggat
     3781 ggctgaggag tactgccaca aacactatga ccgaaacaaa gatggcaaca aagatgtgta
     3841 tctgtccctg cttcggatgt acctgtcgcc ccccagcatt cactgcctgg ggccaatcaa
     3901 gctggaacta ctggagccaa aagccaacct ccaggccgct ctgcaggtcc tcgagctaca
     3961 ccacagcaaa ctggacacca ccaaggccct caaccttctg ccagcaaaca ctcagatcaa
     4021 tgacatacgc atcttcctgg aaaaggtctt ggaagaaaat gcacaaaaga aacggttcaa
     4081 tcaagtgctc aagaaccttc tccatgcaga attcctgagg gtccaggaag agcggatttt
     4141 acaccagcag gtgaagtgca tcatcacaga ggagaaggtg tgcatggtgt gtaagaagaa
     4201 gattgggaac agtgcatttg caagataccc caatggagtg gtcgtccatt acttctgttc
     4261 caaagaggta aacccagctg acactcatca tcaccatcac cattgaggat cctctagagg
     4321 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4381 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4441 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4501 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4561 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     4621 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4681 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4741 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4801 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4861 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4921 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4981 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5041 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5101 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5221 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5281 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5341 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5401 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5461 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5521 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5581 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5641 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5701 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5761 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5821 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5881 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5941 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6001 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6061 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6121 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6181 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6241 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6301 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6361 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6421 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6541 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6601 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6661 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6721 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6781 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6841 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7501 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     7561 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7621 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7861 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7921 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8281 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8581 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8641 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8701 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8761 gagatctccc gatcccctat ggt
//
