LOCUS       V039916                 9273 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039916
VERSION     V039916
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..3953
                     /label="PDZRN3(NM_015009)"
                     /note="PDZRN3(NM_015009)"
                     /gene="PDZRN3"
     CDS             3960..4025
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4049..4637
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4703..4751
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4953..5381
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5395..5724
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5791..6585
                     /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    6761..6882
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6931..6947)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6955..6971
                     /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(6979..7009)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7024..7045
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7333..7921)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8092..8952)
                     /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(8953..9057)
                     /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 ccaccatggg cttcgagctg gaccgcttcg
      781 acggcgacgt ggacccggac ctgaagtgcg cgctgtgcca caaggtcctg gaggacccgc
      841 tgaccacgcc gtgcggccac gtcttctgcg ccggctgcgt gctgccctgg gtggtgcagg
      901 agggcagctg cccggcgcgc tgccgcggtc gcctgtcggc caaagagctc aaccacgtcc
      961 tgccgctcaa gcgccttatc ctcaagctgg acatcaagtg cgcgtacgcg acgcgcggct
     1021 gcggccgggt ggtcaagctg cagcagctgc cggagcacct cgagcgctgc gacttcgcgc
     1081 ccgcgcgctg tcgccacgcg ggttgcggcc aggtgctgct gcggcgcgac gtggaggcgc
     1141 acatgcgcga cgcctgcgac gcgcggccag tgggccgctg ccaggagggc tgcgggctac
     1201 ccttgacgca cggcgagcag cgcgcgggcg gccactgctg cgcgcgagcg ctgcgggcgc
     1261 acaacggcgc gctccaggcc cgcctgggcg cgctgcacaa ggcgctcaag aaggaggcgc
     1321 tgcgcgctgg gaagcgcgag aagtcgctgg tggcccagct ggccgcggcg cagcttgagc
     1381 tgcagatgac cgcgctgcgc taccagaaga aattcaccga atacagcgcg cgcctcgact
     1441 cgctcagccg ctgcgtggcc gcgccgcccg gcggcaaggg cgaagaaacc aaaagtctga
     1501 ctcttgtcct gcatcgggac tccggctccc tgggattcaa tattattggt ggccggccga
     1561 gtgtggataa ccacgatgga tcatccagtg aaggaatctt tgtatccaag atagttgaca
     1621 gtgggcctgc agccaaggaa ggaggcctgc aaattcatga caggattatt gaggtcaacg
     1681 gcagagactt atccagagca actcatgacc aggctgtgga agctttcaag acagccaagg
     1741 agcccatagt ggtgcaggtg ttgagaagaa caccaaggac caaaatgttc acgcctccat
     1801 cagagtctca gctggtggac acgggaaccc aaaccgacat cacctttgaa catatcatgg
     1861 ccctcactaa gatgtcctct cccagcccac ccgtgctgga tccctatctc ttgccagagg
     1921 agcatccctc agcccatgaa tactacgatc caaatgacta cattggagac atccatcagg
     1981 agatggacag ggaggagctg gagctggagg aagtggacct ctacagaatg aacagccagg
     2041 acaagctggg cctcactgtg tgctaccgga cggacgatga agacgacatt gggatttata
     2101 tcagtgagat tgaccctaac agcattgcag ccaaggatgg gcgcatccga gaaggagacc
     2161 gcattatcca gattaatggg atagaggtgc agaaccgtga agaggctgtg gctcttctaa
     2221 ccagtgaaga aaataaaaac ttttcattgc tgattgcaag gcctgaactc cagctggatg
     2281 agggctggat ggatgatgac aggaacgact ttctggatga cctgcacatg gacatgctgg
     2341 aggagcagca ccaccaggcc atgcaattca cagctagcgt gctgcagcag aagaagcacg
     2401 acgaagacgg tgggaccaca gatacagcca ccatcttgtc caaccagcac gagaaggaca
     2461 gcggtgtggg gcggaccgac gagagcaccc gtaatgacga gagctcggag caagagaaca
     2521 atggcgacga cgccaccgca tcctccaacc cgctggcggg gcagaggaag ctcacctgca
     2581 gccaggacac cttgggcagc ggcgacctgc ccttcagcaa cgagtctttc atttcggccg
     2641 actgcacgga cgccgactac ctggggatcc cggtggacga gtgcgagcgc ttccgcgagc
     2701 tcctggagct caagtgccag gtgaagagcg ccacccctta cggcctgtac taccctagcg
     2761 gccccctgga cgccggcaag agtgaccctg agagcgtgga caaggagctg gagctgctga
     2821 acgaagagct gcgcagcatc gagctggagt gcctgagcat cgtgcgcgcc cacaagatgc
     2881 agcagctcaa ggagcagtac cgcgagtcct ggatgctgca caacagcggc ttccgcaact
     2941 acaacaccag catcgacgtg cgcagacacg agctctcaga tatcaccgag ctcccggaga
     3001 aatccgacaa ggacagctcg agcgcctaca acacaggcga gagctgccgc agcaccccgc
     3061 tcaccctgga gatctccccc gacaactcct tgaggagagc ggcggagggc atcagctgcc
     3121 cgagcagcga aggggctgtg gggaccacgg aagcctacgg gccagcctcc aagaatctgc
     3181 tctccatcac ggaagatccc gaagtgggca cccctaccta tagcccgtcc ctgaaggagc
     3241 tggaccccaa ccagcccctg gaaagcaaag agcggagagc cagcgacggg agccggagcc
     3301 ccacgcccag ccagaagctg ggcagcgcct acctgccctc ctatcaccac tccccataca
     3361 agcacgcgca catcccggcg cacgcccagc actaccagag ctacatgcag ctgatccagc
     3421 agaagtcggc cgtggagtac gcgcaaagcc agatgagcct ggtgagcatg tgcaaggacc
     3481 tgagctctcc caccccgtcg gagccgcgca tggagtggaa ggtgaagatc cgcagcgacg
     3541 ggacgcgcta catcaccaag aggcccgtgc gggaccgcct gctgcgggag cgcgccctga
     3601 agatccggga agagcgcagc ggcatgacca ccgacgacga cgcggtgagc gagatgaaga
     3661 tggggcgcta ctggagcaag gaggagagga agcagcacct ggtgaaggcc aaggagcagc
     3721 ggcggcggcg cgagttcatg atgcagagca ggttggattg tctcaaggag cagcaagcag
     3781 ccgatgacag gaaggagatg aacattctcg aactgagcca caaaaagatg atgaagaaga
     3841 ggaataagaa aatcttcgat aactggatga cgatccaaga actcttaacc cacggcacaa
     3901 aatccccgga cggcactaga gtatacaatt ccttcctatc ggtgactact gtaggatccg
     3961 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     4021 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     4081 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4141 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4201 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4261 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4321 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4381 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4441 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4501 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4561 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4621 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     4681 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     4741 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4801 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4861 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     4921 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4981 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5041 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5101 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5161 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5221 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5281 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     5341 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     5401 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5461 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5521 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5581 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5641 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     5701 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     5761 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5821 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5881 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5941 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     6001 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6061 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6121 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6181 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6241 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6301 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6361 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6421 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6481 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6541 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6601 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6661 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6721 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6781 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6841 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6901 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6961 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7021 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7081 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7141 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7201 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7501 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     7561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7741 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8041 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8101 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8161 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8221 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8281 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8341 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8401 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8461 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8521 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8581 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8641 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8701 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8761 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8821 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8881 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8941 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9001 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9061 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     9121 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     9181 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     9241 ttgaccgaca attgcatgaa gaatctgctt agg
//
