LOCUS       V026635                 8381 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026635
VERSION     V026635
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3871
                     /label="Epha2(NM_010139)"
                     /note="Epha2(NM_010139)"
                     /gene="Epha2"
     CDS             3878..3943
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3981..4205
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4251..4679
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4693..5022
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5089..5883
                     /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    6057..6178
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6227..6243)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6251..6267
                     /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(6275..6305)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6320..6341
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6629..7214)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7385..8245)
                     /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(8246..8350)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggagctcc gggcagtcgg
      961 tttctgcctg gcgctgctgt ggggttgcgc gctggcggcc gcggcggcac agggaaagga
     1021 agttgttttg ttggacttcg cagcaatgaa gggagagctc ggctggctca cgcaccccta
     1081 tggcaaaggg tgggacctga tgcagaacat catggacgac atgcctatct acatgtactc
     1141 ggtgtgcaac gtggtatccg gcgaccagga caactggctc cgcaccaact gggtgtaccg
     1201 ggaggaggcc gagcgcatct ttattgagct caagttcacg gtgcgagact gtaacagctt
     1261 cccgggtggc gccagctcat gcaaagagac cttcaacctc tactatgcag agtcagatgt
     1321 ggactatggc accaacttcc agaagcgcca gttcaccaag attgacacca tcgcccctga
     1381 cgagatcacg gtcagcagtg acttcgaggc tcgcaatgtc aagctgaacg tagaggagcg
     1441 catggtgggg ccccttaccc ggaagggctt ctacctggcc ttccaggaca tcggcgcctg
     1501 cgtggcgctg ctctccgttc gcgtctacta caagaagtgt cccgagatgc tgcagagctt
     1561 ggctcgcttc cccgagacca ttgctgtcgc tgtctccgat acacaacccc tggccacggt
     1621 ggccggtacc tgcgtggacc atgccgtggt gccttatggg ggcgaggggc ctctcatgca
     1681 ctgcacggtg gatggcgagt ggctggtgcc catcgggcag tgcctgtgcc aggaaggcta
     1741 cgagaaggtc gaggatgcct gccgagcctg ttctccagga ttcttcaagt ctgaggcatc
     1801 tgagagccct tgcctggagt gtccagagca taccctgcca tccacagagg gtgccacctc
     1861 ctgccagtgt gaagaaggct atttcagggc acctgaggac ccactgtcca tgtcttgcac
     1921 acgtccaccc tctgccccca actacctcac ggccattggc atgggtgcca aagtagaact
     1981 gcgttggaca gctcccaagg acactggtgg ccgccaggac attgtctaca gtgtcacttg
     2041 cgaacagtgc tggccagagt ctggcgagtg tgggccctgt gaggcgagcg tgcgctattc
     2101 agaacctcct cacgccctga cccgcacgag tgtgacagtc agtgacctgg agccccacat
     2161 gaactatacc ttcgctgtcg aagcacgcaa tggtgtctca ggcctggtga ctagccgaag
     2221 cttccggact gccagcgtca gtattaacca aacagagccc cccaaagtga ggctggagga
     2281 ccgaagcacc acctccctga gtgtcacctg gagcatcccg gtgtcacagc agagccgtgt
     2341 gtggaagtac gaagtcacct accgcaagaa gggggatgcc aacagctata atgtgcgccg
     2401 cacggaaggc ttctccgtga ccctggatga ccttgctccg gataccacgt acctggtgca
     2461 ggtgcaggcg ctgacgcagg agggccaggg agccggcagc aaagtgcacg agttccagac
     2521 actgtccacg gaaggatctg ccaacatggc ggtgatcggc ggtgtggctg taggtgttgt
     2581 tttgcttctg gtactggcag gagttggcct cttcatccat cgaaggagga ggaacctgcg
     2641 ggctcgccag tcctctgagg atgtccgttt ttccaagtca gaacaactaa agcccctgaa
     2701 gacctatgtg gatcctcaca cttacgaaga ccccaaccag gctgtactca agtttaccac
     2761 cgagatccac ccatcctgtg tggcaaggca gaaggtcatt ggagcaggag agtttggaga
     2821 ggtctataaa gggacgctga aggcatcctc ggggaagaag gagataccgg tggccatcaa
     2881 gacactgaaa gcgggctaca ctgagaagca gcgggtggac ttcctgagcg aggccagcat
     2941 catgggccag tttagccacc acaatatcat ccgcctggag ggcgtggtct ctaaatacaa
     3001 acccatgatg attatcacag agtacatgga gaatggagcg ctagacaagt tccttaggga
     3061 gaaggatggt gagttcagtg tacttcagtt ggtgggcatg ctgaggggta tcgcatccgg
     3121 catgaagtac ctggccaaca tgaactacgt gcaccgggac ctggccgccc gcaacatcct
     3181 cgtcaacagc aacctggtgt gcaaggtgtc cgattttggc ctgtcgcgtg tgctggaaga
     3241 tgaccccgag gccacctaca ccacaagtgg cggcaagatc cctattcgat ggacagcccc
     3301 agaggccatt tcctaccgca agttcacctc agccagcgat gtgtggagct acggcattgt
     3361 catgtgggaa gtgatgactt atggcgaacg gccctactgg gaactgtcaa accacgaggt
     3421 catgaaagcc atcaacgacg gcttccggct ccctacgccc atggactgcc cttcagccat
     3481 ttaccagctc atgatgcagt gctggcagca agagcgctcc cgccgaccca agtttgccga
     3541 catcgttagc atcctggaca agctcatccg agcccccgac tccctcaaga cgctggctga
     3601 ctttgatccc cgagtgtcca tccggctgcc cagcaccagc ggctcggagg gagtcccctt
     3661 ccgtacggtg tccgagtggc tggagagcat caagatgcaa cagtacacgg aacacttcat
     3721 ggtggctggc tacacggcca tcgagaaggt ggtacagatg tccaacgaag acatcaaaag
     3781 gatcggagtg cgtcttcctg gccaccagaa gcgtattgcc tacagcctgc tgggactcaa
     3841 ggaccaggtc aacacagtgg ggattcctat cctcgaggac tacaaagacc atgacggtga
     3901 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     3961 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4021 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4081 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4141 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4201 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4261 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4321 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4381 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4441 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4501 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4561 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4621 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4681 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4741 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4861 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4921 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4981 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5041 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5101 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5161 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5221 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5281 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5341 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5401 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5461 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5521 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5581 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5641 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5701 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5761 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5821 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5881 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5941 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6001 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6061 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6121 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6181 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6241 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6301 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6361 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6421 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6541 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6601 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6661 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6721 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6781 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6841 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6901 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6961 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7021 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7081 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7141 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7381 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7561 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7801 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8041 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8221 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8341 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
