LOCUS       V037735                 8040 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037735
VERSION     V037735
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             887..2737
                     /label="SYVN1(NM_172230)"
                     /note="SYVN1(NM_172230)"
                     /gene="SYVN1"
     CDS             2744..2809
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2846..3556
                     /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"
     polyA_signal    3607..3831
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3877..4305
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4319..4648
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4715..5509
                     /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    5683..5804
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5853..5869)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5877..5893
                     /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(5901..5931)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5946..5967
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6255..6840)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7011..7871)
                     /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(7872..7976)
                     /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 ggctagcaac ttaaggatcc gccaccatgt tccgcacggc
      901 agtgatgatg gcggccagcc tggcgctgac cggggctgtg gtggctcacg cctactacct
      961 caaacaccag ttctacccca ctgtggtgta cctgaccaag tccagcccca gcatggcagt
     1021 cctgtacatc caggcctttg tccttgtctt ccttctgggc aaggtgatgg gcaaggtgtt
     1081 ctttgggcaa ctgagggcag cagagatgga gcaccttctg gaacgttcct ggtacgccgt
     1141 cacagagact tgtctggcct tcaccgtttt tcgggatgac ttcagccccc gctttgttgc
     1201 actcttcact cttcttctct tcctcaaatg tttccactgg ctggctgagg accgtgtgga
     1261 ctttatggaa cgcagcccca acatctcctg gctctttcac tgccgcattg tctctcttat
     1321 gttcctcctg ggcatcctgg acttcctctt cgtcagccac gcctatcaca gcatcctgac
     1381 ccgtggggcc tctgtgcagc tggtgtttgg ctttgagtat gccatcctga tgacgatggt
     1441 gctcaccatc ttcatcaagt atgtgctgca ctccgtggac ctccagagtg agaacccctg
     1501 ggacaacaag gctgtgtaca tgctctacac agagctgttt acaggcttca tcaaggttct
     1561 gctgtacatg gccttcatga ccatcatgat caaggtgcac accttcccac tctttgccat
     1621 ccggcccatg tacctggcca tgagacagtt caagaaagct gtgacagatg ccatcatgtc
     1681 tcgccgagcc atccgcaaca tgaacaccct gtatccagat gccaccccag aggagctcca
     1741 ggcaatggac aatgtctgca tcatctgccg agaagagatg gtgactggtg ccaagagact
     1801 gccctgcaac cacattttcc ataccagctg cctgcgctcc tggttccagc ggcagcagac
     1861 ctgccccacc tgccgtatgg atgtccttcg tgcatcgctg ccagcgcagt caccaccacc
     1921 cccggagcct gcggatcagg ggccaccccc tgccccccac cccccaccac tcttgcctca
     1981 gccccccaac ttcccccagg gcctcctgcc tccttttcct ccaggcatgt tcccactgtg
     2041 gccccccatg ggcccctttc cacctgtccc gcctcccccc agctcaggag aggctgtggc
     2101 tcctccatcc accagtgcag cagccctttc tcggcccagt ggagcagcta caaccacagc
     2161 tgctggcacc agtgctactg ctgcttctgc cacagcatct ggcccaggct ctggctctgc
     2221 cccagaggct ggccctgccc ctggtttccc cttccctcct ccctggatgg gtatgcccct
     2281 gcctccaccc tttgccttcc ccccaatgcc tgtgccccct gcgggctttg ctgggctgac
     2341 cccagaggag ctacgagctc tggagggcca tgagcggcag cacctggagg cccggctgca
     2401 gagcctgcgt aacatccaca cactgctgga cgccgccatg ctgcagatca accagtacct
     2461 caccgtgctg gcctccttgg ggcccccccg gcctgccact tcagtcaact ccactgagga
     2521 gactgccact acagttgttg ctgctgcctc ctccaccagc atccctagct cagaggccac
     2581 gaccccaacc ccaggagcct ccccaccagc ccctgaaatg gaaaggcctc cagctcctga
     2641 gtcagtgggc acagaggaga tgcctgagga tggagagccc gatgcagcag agctccgccg
     2701 gcgccgcctg cagaagctgg agtctcctgt tgcccacctc gaggactaca aagaccatga
     2761 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagg aattcggtgg
     2821 aggcggatca ggaggcggtg gatctatggt gagcaagggc gaggaggata acatggccat
     2881 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     2941 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     3001 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     3061 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     3121 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     3181 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     3241 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     3301 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     3361 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     3421 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     3481 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     3541 cgagctgtac aagtaggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag
     3601 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3661 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3721 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3781 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3841 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3901 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3961 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4021 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4081 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4141 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4201 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4261 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4321 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4381 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4441 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4501 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4561 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4621 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4681 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4741 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4801 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4861 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4921 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4981 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5041 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5101 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5161 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5221 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5281 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5341 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5401 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5461 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5521 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5581 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5641 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5701 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5761 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5821 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5881 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5941 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6001 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6061 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6121 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6181 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6241 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6301 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6361 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6421 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6481 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6541 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6601 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6661 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6721 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6781 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6841 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6901 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6961 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7021 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7081 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7141 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7201 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7261 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7321 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7381 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7441 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7501 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7561 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7621 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7681 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7741 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7801 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7861 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7921 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7981 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
