LOCUS       sequence-173859-        7491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector containing a C-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7491)
  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 7491)
  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-C-FLAG
FEATURES             Location/Qualifiers
     source          1..7491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..24
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(44..63)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(45..63)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(74..91)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(74..90)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(83..105)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    495..629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(532..551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     586..605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      755..1210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(842..861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1052..1073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1327..1684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1327..1518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1350..1370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1535..1670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1735..2529
                     /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(1789..1808)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2399..2418
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2593..2641
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2682..2701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2801..2823
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2947..3051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3052..3120,3121..3912)
                     /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(3270..3289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4083..4671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4572..4591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4825..4842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(4916..5020)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5130..5433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5434..5637
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5588..5608
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5634..5658
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5759..5776
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    5786..5910
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(5935..5940,5941..5958,5959..5965)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6019..6678
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6085..6104)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             7020..7325
                     /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     7204..7223
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(7366..7490)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gactacaaag acgatgacga caagtgagac gtcatagctc tctccctata gtgagtcgta
       61 ttataagcta ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac tgctagcttg
      121 ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
      181 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactagctg
      241 catatgcttg ctgcttgaga gttttgctta ctgagtatga tttatgaaaa tattatacac
      301 aggagctagt gattctaatt gtttgtgtat tttagattca cagtcccaag gctcatttca
      361 ggcccctcag tcctcacagt ctgttcatga tcataatcag ccataccaca tttgtagagg
      421 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
      481 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      541 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      601 tcatcaatgt atcttatcat gtctggatcg atcctgcatt aatgaatcgg ccaacgcgcg
      661 gggagaggcg gtttgcgtat tggctggcgt aatagcgaag aggcccgcac cgatcgccct
      721 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
      781 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
      841 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
      901 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
      961 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1021 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1081 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1141 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1201 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1261 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     1321 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     1381 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     1441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1501 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     1561 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     1621 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     1681 caaaaagctt gattcttctg acacaacagt ctcgaactta aggctagagc caccatgatt
     1741 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     1801 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     1861 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     1921 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     1981 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2041 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2101 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2161 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2221 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     2281 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     2341 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     2401 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     2461 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     2521 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     2581 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     2641 gaatcgatag cgataaggat ccgcgtatgg tgcactctca gtacaatctg ctctgatgcc
     2701 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2761 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2821 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     2881 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2941 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3001 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3061 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3121 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3181 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3241 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3301 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3361 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3421 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3481 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3541 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3601 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3661 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3721 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3781 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3841 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3901 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3961 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4021 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4081 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4141 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4201 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4261 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4321 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4381 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4441 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4501 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4561 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4621 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4681 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4741 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4801 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4861 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagagc ttgcaattcg
     4921 cgctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4981 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5041 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatta
     5101 cgaggccctt tcacgcatta gatgcatgtc gttacataac ttacggtaaa tggcccgcct
     5161 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5221 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     5281 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     5341 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     5401 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     5461 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     5521 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     5581 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     5641 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
     5701 caccgggacc gatccagcct ccggactcta gcctaggccg cggaccatgg cgtactacca
     5761 tcaccatcac catcactcta gatcaacaag tttgtacaaa aaagctgaac gagaaacgta
     5821 aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata
     5881 ctgtaaaaca caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca
     5941 ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca
     6001 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     6061 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     6121 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     6181 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
     6241 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     6301 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     6361 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     6421 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     6481 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     6541 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     6601 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     6661 tggcagggcg gggcgtaaag atctggatcc ggcttactaa aagccagata acagtatgcg
     6721 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     6781 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     6841 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     6901 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     6961 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
     7021 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     7081 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     7141 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     7201 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     7261 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     7321 tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata
     7381 tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg
     7441 atatttatat cattttacgt ttctcgttca gctttcttgt acaaagtggt t
//
