LOCUS       V037707                 8670 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037707
VERSION     V037707
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             744..1451
                     /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             1500..3350
                     /label="SYVN1(NM_172230)"
                     /note="SYVN1(NM_172230)"
                     /gene="SYVN1"
     CDS             3357..3422
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3446..4034
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4100..4148
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4350..4778
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4792..5121
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5188..5982
                     /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    6158..6279
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6328..6344)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6352..6368
                     /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(6376..6406)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6421..6442
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6730..7318)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7489..8349)
                     /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(8350..8454)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca
      781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca
     1501 tgttccgcac ggcagtgatg atggcggcca gcctggcgct gaccggggct gtggtggctc
     1561 acgcctacta cctcaaacac cagttctacc ccactgtggt gtacctgacc aagtccagcc
     1621 ccagcatggc agtcctgtac atccaggcct ttgtccttgt cttccttctg ggcaaggtga
     1681 tgggcaaggt gttctttggg caactgaggg cagcagagat ggagcacctt ctggaacgtt
     1741 cctggtacgc cgtcacagag acttgtctgg ccttcaccgt ttttcgggat gacttcagcc
     1801 cccgctttgt tgcactcttc actcttcttc tcttcctcaa atgtttccac tggctggctg
     1861 aggaccgtgt ggactttatg gaacgcagcc ccaacatctc ctggctcttt cactgccgca
     1921 ttgtctctct tatgttcctc ctgggcatcc tggacttcct cttcgtcagc cacgcctatc
     1981 acagcatcct gacccgtggg gcctctgtgc agctggtgtt tggctttgag tatgccatcc
     2041 tgatgacgat ggtgctcacc atcttcatca agtatgtgct gcactccgtg gacctccaga
     2101 gtgagaaccc ctgggacaac aaggctgtgt acatgctcta cacagagctg tttacaggct
     2161 tcatcaaggt tctgctgtac atggccttca tgaccatcat gatcaaggtg cacaccttcc
     2221 cactctttgc catccggccc atgtacctgg ccatgagaca gttcaagaaa gctgtgacag
     2281 atgccatcat gtctcgccga gccatccgca acatgaacac cctgtatcca gatgccaccc
     2341 cagaggagct ccaggcaatg gacaatgtct gcatcatctg ccgagaagag atggtgactg
     2401 gtgccaagag actgccctgc aaccacattt tccataccag ctgcctgcgc tcctggttcc
     2461 agcggcagca gacctgcccc acctgccgta tggatgtcct tcgtgcatcg ctgccagcgc
     2521 agtcaccacc acccccggag cctgcggatc aggggccacc ccctgccccc caccccccac
     2581 cactcttgcc tcagcccccc aacttccccc agggcctcct gcctcctttt cctccaggca
     2641 tgttcccact gtggcccccc atgggcccct ttccacctgt cccgcctccc cccagctcag
     2701 gagaggctgt ggctcctcca tccaccagtg cagcagccct ttctcggccc agtggagcag
     2761 ctacaaccac agctgctggc accagtgcta ctgctgcttc tgccacagca tctggcccag
     2821 gctctggctc tgccccagag gctggccctg cccctggttt ccccttccct cctccctgga
     2881 tgggtatgcc cctgcctcca ccctttgcct tccccccaat gcctgtgccc cctgcgggct
     2941 ttgctgggct gaccccagag gagctacgag ctctggaggg ccatgagcgg cagcacctgg
     3001 aggcccggct gcagagcctg cgtaacatcc acacactgct ggacgccgcc atgctgcaga
     3061 tcaaccagta cctcaccgtg ctggcctcct tggggccccc ccggcctgcc acttcagtca
     3121 actccactga ggagactgcc actacagttg ttgctgctgc ctcctccacc agcatcccta
     3181 gctcagaggc cacgacccca accccaggag cctccccacc agcccctgaa atggaaaggc
     3241 ctccagctcc tgagtcagtg ggcacagagg agatgcctga ggatggagag cccgatgcag
     3301 cagagctccg ccggcgccgc ctgcagaagc tggagtctcc tgttgcccac ctcgaggact
     3361 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     3421 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     3481 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3541 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3601 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3661 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3721 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3781 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3841 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3901 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3961 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4021 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     4081 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     4141 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     4201 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     4261 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     4321 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4501 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4561 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4621 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4681 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     4741 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4801 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4861 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4921 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4981 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5041 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     5101 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     5161 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5221 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5281 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5341 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5401 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5461 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5521 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5581 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5641 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5701 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5761 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5821 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5881 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5941 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6001 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6061 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6121 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6301 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6361 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6421 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6481 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6901 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     6961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7141 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7501 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8341 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8461 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     8521 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     8581 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     8641 accgacaatt gcatgaagaa tctgcttagg
//
