LOCUS       V026911                 9357 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026911
VERSION     V026911
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..4037
                     /label="SPINK5(NM_001127698)"
                     /note="SPINK5(NM_001127698)"
                     /gene="SPINK5"
     CDS             4044..4109
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4133..4721
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4787..4835
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5037..5465
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5479..5808
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5875..6669
                     /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    6845..6966
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7015..7031)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7039..7055
                     /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(7063..7093)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7108..7129
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7417..8005)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8176..9036)
                     /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(9037..9141)
                     /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 ccaccatgaa gatagccaca gtgtcagtgc
      781 ttctgccctt ggctctttgc ctcatacaag atgctgccag taagaatgaa gatcaggaaa
      841 tgtgccatga atttcaggca tttatgaaaa atggaaaact gttctgtccc caggataaga
      901 aattttttca aagtcttgat ggaataatgt tcatcaataa atgtgccacg tgcaaaatga
      961 tactggaaaa agaagcaaaa tcacagaaga gggccaggca tttagcaaga gctcccaagg
     1021 ctactgcccc aacagagctg aattgtgatg attttaaaaa aggagaaaga gatggggatt
     1081 ttatctgtcc tgattattat gaagctgttt gtggcacaga tgggaaaaca tatgacaaca
     1141 gatgtgcact gtgtgctgag aatgcgaaaa ccgggtccca aattggtgta aaaagtgaag
     1201 gggaatgtaa gagcagtaat ccagagcagg atgtatgcag tgcttttcgg ccctttgtta
     1261 gagatggaag acttggatgc acaagggaaa atgatcctgt tcttggtcct gatgggaaga
     1321 cgcatggcaa taagtgtgca atgtgtgctg agctgttttt aaaagaagct gaaaatgcca
     1381 agcgagaggg tgaaactaga attcgacgaa atgctgaaaa ggatttttgc aaggaatatg
     1441 aaaaacaagt gagaaatgga aggctttttt gtacacggga gagtgatcca gtccgtggcc
     1501 ctgacggcag gatgcatggc aacaaatgtg ccctgtgtgc tgaaattttc aagcagcgtt
     1561 tttcagagga aaacagtaaa acagatcaaa atttgggaaa agctgaagaa aaaactaaag
     1621 ttaaaagaga aattgtgaaa ctctgcagtc aatatcaaaa tcaggcaaag aatggaatac
     1681 ttttctgtac cagagaaaat gaccctattc gtggtccaga tgggaaaatg catggcaact
     1741 tgtgttccat gtgtcaagcc tacttccaag cagaaaatga agaaaagaaa aaggctgaag
     1801 cacgagctag aaacaaaaga gaatctggaa aagcaacctc atatgcagag ctttgcagtg
     1861 aatatcgaaa gcttgtgagg aacggaaaac ttgcttgcac cagagagaac gatcctatcc
     1921 agggcccaga tgggaaagtg catggcaaca cctgctccat gtgtgaggtc ttcttccaag
     1981 cagaagaaga agaaaagaaa aagaaggaag gtaaatcaag aaacaaaaga caatctaaga
     2041 gtacagcttc ctttgaggag ttgtgtagtg aataccgcaa atccaggaaa aacggacggc
     2101 ttttttgcac cagagagaat gaccccatcc agggcccaga tggaaaaatg catggcaaca
     2161 cctgctccat gtgtgaggcc ttctttcaac aagaagaaag agcaagagca aaggctaaaa
     2221 gagaagctgc aaaggaaatc tgcagtgaat ttcgggacca agtgaggaat ggaacactta
     2281 tatgcaccag ggagcataat cctgtccgtg gcccagatgg caaaatgcat ggaaacaagt
     2341 gtgccatgtg tgccagtgtg ttcaaacttg aagaagaaga gaagaaaaat gataaagaag
     2401 aaaaagggaa agtcgaggct gaaaaagtta agagagaagc agttcaggag ctgtgcagtg
     2461 aatatcgtca ttatgtgagg aatggacgac tcccctgtac cagagagaat gatcctattg
     2521 agggtctaga tgggaaaatc cacggcaaca cctgctccat gtgtgaagcc ttcttccagc
     2581 aagaagcaaa agaaaaagaa agagctgaac ccagagcaaa agtcaaaaga gaagctgaaa
     2641 aggagacatg cgatgaattt cggagacttt tgcaaaatgg aaaacttttc tgcacaagag
     2701 aaaatgatcc tgtgcgtggc ccagatggca agacccatgg caacaagtgt gccatgtgta
     2761 aggcagtctt ccagaaagaa aatgaggaaa gaaagaggaa agaagaggaa gatcagagaa
     2821 atgctgcagg acatggttcc agtggtggtg gaggaggaaa cactcaggac gaatgtgctg
     2881 agtatcggga acaaatgaaa aatggaagac tcagctgtac tcgggagagt gatcctgtac
     2941 gtgatgctga tggcaaatcg tacaacaatc agtgtaccat gtgtaaagca aaattggaaa
     3001 gagaagcaga gagaaaaaat gagtattctc gctccagatc aaatgggact ggatcagaat
     3061 cagggaagga tacatgtgat gagtttagaa gccaaatgaa aaatggaaaa ctcatctgca
     3121 ctcgagaaag tgaccctgtc cggggtccag atggcaagac acatggcaat aagtgtacta
     3181 tgtgtaagga aaaactggaa agggaagcag ctgaaaaaaa aaagaaagag gatgaagaca
     3241 ggagcaatac aggagaaagg agcaatacag gagaaaggag caatgacaaa gaggatctgt
     3301 gtcgtgaatt tcgaagcatg cagagaaatg gaaagcttat ctgcaccaga gaaaataacc
     3361 ctgttcgagg cccatatggc aagatgcaca tcaataaatg tgctatgtgt cagagcatct
     3421 ttgatcgaga agctaatgaa agaaaaaaga aagatgaaga gaaatcaagt agcaagccct
     3481 caaataatgc aaaggaccag tgcagacagg ttcagaatga agcggaggat gcaaaattta
     3541 gacaacctgg gcgttccttg gcctctgttg ccaggatgag tacagatgag tgcagtgaat
     3601 ttcgaaacta tataaggaac aatgaactca tctgccctag agagaatgac ccagtgcacg
     3661 gtgctgatgg aaagttctat acaaacaagt gctacatgtg cagagctgtc tttctaacag
     3721 aagctttgga aagggcaaag cttcaagaaa agccatccca tgttagagct tctcaagagg
     3781 aagacagccc agactctttc agttctctgg attctgagat gtgcaaagac taccgagtat
     3841 tgcccaggat aggttatctt tgtccaaagg atttaaagcc tgtctgtggt gacgatggcc
     3901 aaacctacaa caatccttgc atgctctgtc atgaaaacct gatacgccaa acaaatacac
     3961 acatccgcag tacagggaag tgtgaggaga gcagcacccc aggaaccacc gcagccagca
     4021 tgcccccgtc tgacgaagga tccgactaca aagaccatga cggtgattat aaagatcatg
     4081 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     4141 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4201 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4261 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     4321 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4381 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4441 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4501 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4561 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4621 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4681 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     4741 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4801 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4861 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4921 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4981 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     5041 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5101 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5161 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     5221 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5281 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5341 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5401 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5461 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     5761 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     5821 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5881 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5941 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6001 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     6061 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6121 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6181 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6241 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6301 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6361 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     6421 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6481 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6541 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6601 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6661 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     6721 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6781 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6841 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6901 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6961 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     7021 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7081 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7141 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7201 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7261 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     7621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8941 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9001 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9061 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9121 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     9181 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     9241 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     9301 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
