LOCUS       V036304                 9189 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036304
VERSION     V036304
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..3869
                     /label="PITRM1(NM_001242307)"
                     /note="PITRM1(NM_001242307)"
                     /gene="PITRM1"
     CDS             3876..3941
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3965..4553
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4619..4667
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4869..5297
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5311..5640
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5707..6501
                     /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    6677..6798
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6847..6863)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6871..6887
                     /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(6895..6925)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6940..6961
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7249..7837)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8008..8868)
                     /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(8869..8973)
                     /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 ccaccatgtg gcgctgcggc gggcggcagg
      781 gcctgtgtgt gctgaggcgg ctgagcggcg gacatgcaca ccacagagcg tggcgatgga
      841 acagtaaccg ggcttgtgag agggctctgc agtataaact aggagacaag atccatggat
      901 tcaccgtaaa ccaggtgaca tctgttcccg agctgttcct gactgcagtg aagctcaccc
      961 atgatgacac aggagccagg tatttacacc tggccagaga agacacgaat aatctgttca
     1021 gcgtgcagtt ccgtactact cccatggaca gtactggtgt tcctcacatt cttgagcata
     1081 ccgtcctttg tgggtctcag aaatatccgt gcagagaccc tttcttcaaa atgttgaacc
     1141 ggtccctctc cacgttcatg aacgccttca cagctagtga ttatactctg tatccatttt
     1201 ccacacaaaa tcccaaggac tttcagaatc tcctctcggt gtatttggat gccacctttt
     1261 tcccatgttt acgcgagctg gatttctggc aggaaggatg gcggctggaa catgagaatc
     1321 cgagcgaccc ccagacgccc ttggtcttta aaggagtcgt ctttaatgag atgaagggag
     1381 cgtttacaga caatgagagg atattctccc agcaccttca gaacagactt cttcctgacc
     1441 acacgtactc agtggtctcc gggggtgacc cactgtgcat cccggagctt acatgggagc
     1501 agcttaagca gtttcatgcc actcactatc acccaagcaa tgctaggttc ttcacgtacg
     1561 gtaattttcc attagaacag catctgaaac aaattcacga ggaagcactg agcaaattcc
     1621 agaaaattga accaagcacc gtggtgccag ctcagacacc ctgggacaag cctagggaat
     1681 tccagataac atgtggcccg gattcatttg ctacagatcc ctctaaacaa acaaccatca
     1741 gcgttagctt cctcttaccg gacatcaccg acacatttga agccttcaca ttaagtcttc
     1801 tgtcttcact cttgacttct gggcccaatt ctccctttta caaagccttg attgaatctg
     1861 gccttggcac agacttttct cctgatgttg gatataatgg ctacacgagg gaggcctact
     1921 ttagtgtcgg cctccaaggg attgcggaga aagacattga gaccgtcaga agcctcatag
     1981 acagaacgat tgatgaagta gttgagaaag gatttgaaga tgatcgaatt gaggctttac
     2041 ttcataaaat tgaaatacag atgaaacatc agtctaccag ctttgggctg atgctgacat
     2101 catacatagc ttcttgctgg aaccatgatg gggaccctgt ggagctcttg aagttgggaa
     2161 atcagttagc taaattcaga cagtgcctgc aggaaaatcc aaaatttttg caagaaaaag
     2221 taaaacagta ttttaagaat aaccagcata agctgacttt atcgatgagg ccagatgaca
     2281 agtatcacga gaagcaggca caggtggaag ccacgaagct caagcagaag gtcgaggctc
     2341 tgtcccccgg agacaggcag cagatctacg agaaaggtct agaattacgg agtcaacaaa
     2401 gcaaacctca agatgcctct tgtctgccag cgttgaaagt ttccgatatt gaacccacca
     2461 tacctgtcac agagttggac gtggtcctga cagctggaga tatccctgtt cagtactgcg
     2521 cccagcccac caatggcatg gtgtatttcc gggccttctc cagcctgaac acactccccg
     2581 aggagctgag gccctatgtg cccctcttct gcagcgtcct caccaagctg ggctgcggcc
     2641 ttcttgacta ccgggagcag gctcagcaga tagaattgaa gaccggaggg atgagtgctt
     2701 ctccccacgt gctccccgac gactcacaca tggacaccta cgagcaggta ggtgtgcttt
     2761 tctcctctct ctgcctggat cgaaacctgc cagacatgat gcagctatgg agtgaaatat
     2821 ttaacaaccc gtgctttgaa gaagaggagc acttcaaggt gctggtgaag atgaccgccc
     2881 aggagctcgc caatggaatt cctgactctg ggcacctgta cgcatccatc agggcaggcc
     2941 ggaccctcac gcccgcaggg gacctgcagg agaccttcag cgggatggat caggtgcggc
     3001 tgatgaagag gattgcagaa atgacagata tcaaacccat cctgaggaag ctcccgcgta
     3061 tcaagaaaca cttgttaaat ggtgataata tgaggtgttc agtgaatgcg actcctcagc
     3121 agatgcctca gacagaaaaa gcggtcgaag acttccttag aagcatcggt cggagtaaaa
     3181 aggaacggag gcctgtgcgc ccacacacgg tcgagaaacc tgtgcccagc agctctggtg
     3241 gagatgccca cgttccccat ggctcccagg tcattaggaa gctggtcatg gaacccacct
     3301 tcaagccctg gcagatgaag actcacttcc tgatgccctt cccggtgaat tacgtgggtg
     3361 aatgcatccg aactgtcccc tacacggacc cagatcatgc cagtcttaaa atccttgcac
     3421 gtttgatgac tgccaaattc ttgcatacag aaattcgaga aaaaggcggt gcttatggtg
     3481 gaggcgcaaa actcagccac aatgggattt tcacccttta ctcttacagg gacccaaata
     3541 caatagagac gctccagtct tttgggaagg ctgtcgactg ggctaagtct ggaaaattca
     3601 cacagcaaga catcgacgaa gccaaacttt ctgtcttctc aaccgtagat gctcctgtcg
     3661 ctccttcaga caaaggaatg gaccacttct tgtacggcct ctcggatgag atgaagcagg
     3721 cccacagaga gcagctcttt gctgtcagcc acgacaagct cctggccgtg agcgatagat
     3781 acctcggcac tgggaagagc acacacggcc tggccatcct cggacccgag aacccgaaaa
     3841 ttgccaagga cccatcctgg atcatccaag gatccgacta caaagaccat gacggtgatt
     3901 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     3961 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4021 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4081 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4141 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4201 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4261 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4321 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4381 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4441 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4501 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     4561 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4621 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     4681 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4741 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4801 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     4861 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4921 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4981 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     5041 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5101 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5161 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5221 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     5281 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5341 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5401 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5461 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5521 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5581 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5641 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5701 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5761 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5821 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5881 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5941 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6001 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6061 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6121 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6181 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6241 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6301 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6361 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6421 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6481 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     6541 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6601 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6661 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6721 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6781 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6841 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6901 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6961 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7021 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7081 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7141 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7201 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7261 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7321 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7381 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7441 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7501 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7561 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7621 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7681 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7741 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7801 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7861 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7921 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7981 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8041 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8101 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8161 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8221 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8281 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8341 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8401 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8461 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8521 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8581 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8641 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8701 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8761 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8821 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8881 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8941 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9001 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     9061 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     9121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     9181 ctgcttagg
//
