LOCUS       V047450                 8037 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047450
VERSION     V047450
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             756..2717
                     /label="Syt16(NM_001364281)"
                     /note="Syt16(NM_001364281)"
                     /gene="Syt16"
     CDS             2724..2789
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2813..3401
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3467..3515
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3717..4145
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4159..4488
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4555..5349
                     /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    5525..5646
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5695..5711)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5719..5735
                     /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(5743..5773)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5788..5809
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6097..6685)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6856..7716)
                     /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(7717..7821)
                     /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 tggtaccgag ctcgaattcg ccaccatggt gttgaccatg gcatctcagg
      781 atgtccagaa cttcttccag cctttgtctt cctggctttc ccgagtttat gaagctctcc
      841 agcaagccgg ggatgcatta tctgcttcac tggtccttct tagcaaacac gattcggcat
      901 tgagtgataa gccagagcag gacttagatg ctgcccagat tcagcaaacc tacttggaag
      961 atgaagaaca ggaccacgat ggcagtccag aggaggcaag ctccctgttt cttgaagagg
     1021 accatttctc actctctaat agtgatctgc aggactccgt ccaaacggca agccccacgc
     1081 ttggtcagca agccgaagac tcaagctctg taataccccc gtggcccagt aagatcccag
     1141 gggccccgaa gccccagcct gtgctttctt cgatcgccga ggaagaccat cactctgaga
     1201 ggcagagatg tggccgtcag catggctctg gcactttaga aaaggttaat ggaagaaagc
     1261 aagttaacag ctttggggat gatgaagagc catccacatc ttctgagagt gatgaagatg
     1321 tgactaagca attcaagatc tcagtgtccc gatcccagag ttttcgttca ggagtatctg
     1381 agaagggaaa gacaacagag ttggagcaga aaataaaatg caaacgtttg ctgtgtaccc
     1441 accaggaaga cagcgctgaa gggtctgctt gtgaaggtat tcattgcagt gcattgcatt
     1501 gcaggcaagg gcaggtcaat gatttggatc gaaccagcca acttagttat tctgagattc
     1561 tgtcttatga agaccgcccc atttccattc tcccacaatc ccccttcgag agcaggaacg
     1621 tcaggcacca tggcccttgc cgtccggaga tgggcatggt ccgaagcctc ggtcgtcctt
     1681 gtgcagatgg cgtcctggag acggagacag cttttgtgag tcggggcttt gaagattcct
     1741 atgccaccca cagttcctct ctgtggagtc cagaggagca ggatggcacc agtctgcagg
     1801 taccacacag gcttttggag cctatctcaa aatgcggtga cctcgatgtc atctttgaat
     1861 atagagctgt aacccagaag ctcactgtga ccattgtgag agcacagggt ctcccagata
     1921 aggaccgaag tggtgtcaac tcttggcaag ttcacatagt gctgctgccc agcaagaaac
     1981 agaggggtaa gacgaacata cagagagggc ccaaccctgt cttcaaggaa aaggtcacct
     2041 tcaccaagct ggagcccaga gatgtggctt cctgtgctgt ccgcttccgc ctgtatgctg
     2101 cccgtaagat gacccgggag agaatgatgg gggagaagct gttctgtctc agccacttgc
     2161 acccagaagg ggaaatgaaa gtgactctgg ttctggagcc aagaagtaat ctgagcagtg
     2221 gagagtctcc gctcagccca tctgtggtgt ctcacagtga cagtgcttca tccacgcagt
     2281 cactgtctca tggaggggtg ccagagctgc tggtgggact gtcctacaat gccacaacag
     2341 ggcgattatc tgtggaaatg atcaaaggca gccatttccg caacctcgct gctaaccgag
     2401 ctcctgacac atatggaaaa ctcttcctcc tcaactgtgt gggtcaagag atgtcccgct
     2461 gcaagacgtc cattcgtcgt ggccaaccca atcctgtcta taaagagacc tttgtcttcc
     2521 aggtggccct cttccagctc tcggacgtca ccttgatgat ttccatttac agcaggcgca
     2581 ctatgaagcg taaagagatg atcggctggg tcgctctggg ccagaacagc agtggggagg
     2641 aggagcagga acactgggag gaaatgaagg aaagcaaggg ccagcagacc tgccggtggc
     2701 atacgctgtt ggaatccgga tccgactaca aagaccatga cggtgattat aaagatcatg
     2761 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     2821 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2881 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2941 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3001 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3061 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3121 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3181 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3241 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3301 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3361 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     3421 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     3481 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3541 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3601 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3661 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     3721 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3781 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3841 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     3901 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3961 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4021 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4081 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4141 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4201 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4261 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4321 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4381 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4441 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4501 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4561 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4621 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4681 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4741 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4801 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4861 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4921 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4981 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5041 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5101 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5161 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5221 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5281 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5341 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     5401 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5461 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5521 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5581 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5641 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5701 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5761 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5821 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     6301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6601 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6781 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6841 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7441 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7741 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7801 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7861 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     7921 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7981 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
