LOCUS       sequence-173852-        7474 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination vector containing an N-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7474)
  AUTHORS   Amr S, Heisey C, Zhang M, Xia XJ, Shows KH, Ajlouni K, Pandya A, 
            Satin LS, El-Shanti H, Shiang R
  TITLE     A homozygous mutation in a novel zinc-finger protein, ERIS, is 
            responsible for Wolfram syndrome 2.
  JOURNAL   Am J Hum Genet. 2007 Oct;81(4):673-83. Epub 2007 Aug 20.
  PUBMED    17846994
REFERENCE   2  (bases 1 to 7474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Am J Hum 
            Genet."; date: "2007-10"; volume: "81(4)"; pages: "673-83. Epub 2007
            Aug 20"
COMMENT     Sequence Label: pDEST26-N-FLAG
FEATURES             Location/Qualifiers
     source          1..7474
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..24
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    34..158
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(183..188,189..206,207..213)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             267..926
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(333..352)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1268..1573
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1452..1471
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1614..1738)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1770..1789)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1771..1789)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1800..1817)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1800..1816)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1809..1831)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2221..2355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2258..2277)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2312..2331
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2558..2577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2768..2789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3043..3400
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3043..3234
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3066..3086)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3251..3386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3451..4245
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3505..3524)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4115..4134
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4309..4357
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4398..4417)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4517..4539
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4663..4767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4768..4836,4837..5628)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4986..5005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5799..6387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6288..6307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6541..6558
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6632..6736)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6846..7149
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7150..7353
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7304..7324
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7350..7374
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gactacaaag acgatgacga caagtctaga tcaacaagtt tgtacaaaaa agctgaacga
       61 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
      121 acataatact gtaaaacaca acatatccag tcactatggc ggccgcatta ggcaccccag
      181 gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccgtcga
      241 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
      301 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
      361 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
      421 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
      481 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
      541 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
      601 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
      661 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
      721 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
      781 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
      841 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
      901 gcgatgagtg gcagggcggg gcgtaaagat ctggatccgg cttactaaaa gccagataac
      961 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     1021 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     1081 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     1141 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     1201 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     1261 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     1321 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
     1381 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     1441 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     1501 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     1561 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gaccatagtg
     1621 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     1681 atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggttg
     1741 atcgcgtgca tgcgacgtca tagctctctc cctatagtga gtcgtattat aagctaggca
     1801 ctggccgtcg ttttacaacg tcgtgactgg gaaaactgct agcttgggat ctttgtgaag
     1861 gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct
     1921 aaggtaaata taaaattttt aagtgtataa tgtgttaaac tagctgcata tgcttgctgc
     1981 ttgagagttt tgcttactga gtatgattta tgaaaatatt atacacagga gctagtgatt
     2041 ctaattgttt gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct
     2101 cacagtctgt tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2161 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2221 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2281 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2341 tatcatgtct ggatcgatcc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2401 gcgtattggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2461 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2521 ggttacgcgc agcgtgaccg ctacacttgc cagcccgctc ctttcgcttt cttcccttcc
     2581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2761 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2821 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2881 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     2941 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     3001 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     3061 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3121 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3181 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3241 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3301 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3361 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac
     3421 aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt
     3481 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3541 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3601 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3661 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3721 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3781 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3841 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3901 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3961 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4021 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4081 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4141 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4201 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4261 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct
     4321 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc
     4381 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4441 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4501 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4561 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4621 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4681 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4741 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4801 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4861 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4921 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4981 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5041 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5101 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5161 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5221 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5281 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5341 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5401 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5461 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5521 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5581 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5641 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5701 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5761 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5821 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5881 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5941 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6001 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6061 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6121 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6181 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6241 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6301 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6361 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6421 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6481 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6541 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6601 cgttggccga ttcattaatg cagagcttgc aattcgcgct ttttcaatat tattgaagca
     6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6781 ttatcatgac attaacctat aaaaataggc gtattacgag gccctttcac gcattagatg
     6841 catgtcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     6901 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     6961 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     7021 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     7081 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     7141 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     7201 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     7261 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     7321 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     7381 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
     7441 actctagcct aggccgcgga ccatggcgta ctac
//
