LOCUS       Exported                6970 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human KLF11.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_KLF11
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  AUTHORS   Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts 
            BS, Mackiewicz M, Mendenhall EM, Myers RM
  TITLE     CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins.
  JOURNAL   Genome Res. 2015 Sep 9.
  PUBMED    26355004
REFERENCE   2  (bases 1 to 6970)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6970
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      27..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     769..786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1086..1107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1122..1152
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1160..1176
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1165..1187
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1184..1200
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1184..1200
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1377..1579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1530..1550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1576..1600
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1676..2806
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     CDS             3840..3863
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3867..3890
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3894..3917
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3933..3989
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4002..4796
                     /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(4056..4075)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4666..4685
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5567..5584)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5567..5583)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5576..5598)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5802..6617)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNILQFHLMLDEFF"
     primer_bind     6525..6544
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6609..6628
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
       61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      781 agcgaggaag cggaagaagc tcgcacattc agcagcgttt ttcagcgcgt tttcgatcaa
      841 cgtttcaatg ttggtatcaa caccaggttt aactttgaac ttatcggcac tgacggttac
      901 tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct
      961 gatccttcaa ctcagcaaaa gttcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1021 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1081 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     1141 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     1201 catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag
     1261 ctcgaattag cttgcatgcc tgcaggtcaa ttccctggca ttatgcccag tacatgacct
     1321 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1381 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttcaag
     1441 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1501 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1561 ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     1621 ctgttttgac ctccatagga gacaccggga ccgatccagc ctggggatcc cgaccatggc
     1681 ttcgtacccc tgccatcaac acgcgtctgc gttcgaccag gctgcgcgtt ctcgcggcca
     1741 tagcaaccga cgtacggcgt tgcgccctcg ccggcagcaa gaagccacgg aagtccgcct
     1801 ggagcagaaa atgcccacgc tactgcgggt ttatatagac ggtcctcacg ggatggggaa
     1861 aaccaccacc acgcaactgc tggtggccct gggttcgcgc gacgatatcg tctacgtacc
     1921 cgagccgatg acttactggc aggtgctggg ggcttccgag acaatcgcga acatctacac
     1981 cacacaacac cgcctcgacc agggtgagat atcggccggg gacgcggcgg tggtaatgac
     2041 aagcgcccag ataacaatgg gcatgcctta tgccgtgacc gacgccgttc tggctcctca
     2101 tatcgggggg gaggctggga gctcacatgc cccgcccccg gccctcaccc tcatcttcga
     2161 ccgccatccc atcgccgccc tcctgtgcta cccggccgcg cgatacctta tgggcagcat
     2221 gaccccccag gccgtgctgg cgttcgtggc cctcatcccg ccgaccttgc ccggcacaaa
     2281 catcgtgttg ggggcccttc cggaggacag acacatcgac cgcctggcca aacgccagcg
     2341 ccccggcgag cggcttgacc tggctatgct ggccgcgatt cgccgcgttt acgggctgct
     2401 tgccaatacg gtgcggtatc tgcagggcgg cgggtcgtgg cgggaggatt ggggacagct
     2461 ttcggggacg gccgtgccgc cccagggtgc cgagccccag agcaacgcgg gcccacgacc
     2521 ccatatcggg gacacgttat ttaccctgtt tcgggccccc gagttgctgg cccccaacgg
     2581 cgacctgtac aacgtgtttg cctgggcctt ggacgtcttg gccaaacgcc tccgtcccat
     2641 gcacgtcttt atcctggatt acgaccaatc gcccgccggc tgccgggacg ccctgctgca
     2701 acttacctcc gggatggtcc agacccacgt caccaccccc ggctccatac cgacgatctg
     2761 cgacctggcg cgcacgtttg cccgggagat gggggaggct aactgaggat cccacgtcac
     2821 tattgtatac tctatattat actctatgtt atactctgta atcctactca ataaacgtgt
     2881 cacgcctgtg aaaccgtact aagtctcccg tgtattctta tcaccatcag gtgacatcct
     2941 cgcccaggct gtcaatcatg ccggtatcga ttccagtagc accggcccca cgctgacaac
     3001 ccactcttgc agcgttagca gcgcccctct taacaagccg acccccacca gcgtcgcggt
     3061 tactaacact cctctccccg acctgcagcc cagctgtgag ccactgcgcc tggctggatg
     3121 ctacattttt aattaattta tttttgtttt ttggtaaaga ttgggtttta ctttgttgcc
     3181 caggctagtc tggaactcct gtgctcaact gatcgtctcc agcctcccaa agtgctgcga
     3241 ttataggttt gagccgctgt acttgtactt cgcctgattt atttttaaat atattttgaa
     3301 agattagctt gaaatctttg ggtgctaaca tagctgcatc ttaagtgtgg gaggaataaa
     3361 tgccttttgg aggatttctt gacccaaatc atccaattcc agagtaataa gataccactt
     3421 tcttctgagt ttgatgaaca cgattttaaa atgacatgaa ttaacccctt gaataaggtg
     3481 cttagcgttt cctttctctt taatatgtat tttctcacct cacaggggag aagcctttca
     3541 actgcagctg ggatggctgt gataaaaagt ttgctcgttc ggatgagctg tcacgccacc
     3601 gcagaactca cacaggggag aagaagtttg tgtgcccggt gtgtgaccga cgtttcatgc
     3661 gcagtgacca cctgacgaag catgcccggc gccacatgac gaccaagaag atcccaggct
     3721 ggcaggcaga ggttggcaag ctgaacagaa tcgcctctgc agagagcccg gggagcccac
     3781 tggtgagcat gccagcctct gcagggagcg gaggaggttc cggtggaggt ggttctggag
     3841 attacaagga tgacgacgat aagggcgatt acaaggatga cgacgataag ggagattaca
     3901 aggatgacga cgataaggtt tcaggaagcg gagctactaa cttcagcctg ctgaagcagg
     3961 ctggagacgt ggaggagaac cctggacctg gatcgtttcg catgattgaa caagatggat
     4021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4501 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4561 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4621 tcgactgtgg ccggctgggt gtggccgacc gctatcagga catagcgttg gctacccgtg
     4681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg
     4801 aacatcatta ggacatcact catgggattt ttaaaaagcc tctttccagg aatggaactg
     4861 atggattcct ctcccactgc ctcacccaaa aaaaacggtc ttggcggcct aggggaagat
     4921 cggggagctg gttttgatga aagtatgtta acttttcttt tccacttggg accctgttca
     4981 gtatcttttg tagtttcaga agtttttttg ttttggtttt ttttttaaag aaatggtaga
     5041 aaatttgata atctgaatca ccagcattca aacaaatatt tcggcaataa agtttacaaa
     5101 atctggattt ttacaacctt ttctattgat gttttgtaga aataagacag ggtactaatt
     5161 tttatactgg tttttagaaa aatatttata ttgttggtgc tcaaatcacc aatttctagc
     5221 tagatcattt tgcagccttc ttttcagtgt ttaataacaa agtttttcct aatggccctt
     5281 cttttagtaa actggacatg ttattccact acaaaaacca caagttatct ggccttttag
     5341 atctttttgg aatcggacct ggttgagtaa ggacctctta aaagggaaaa ataaattttg
     5401 ccgtcagctt cttcataacg ttttcaagga aattctaggc aatcattcct gtcaccaaag
     5461 aactaaaatt ttggttgact ggaactagtg agctgtgtcc atggtgtgtc atgacttgga
     5521 aagtcctctc cactgactgt agcctccaat tcactggccg tcgttcactg gccgtcgttt
     5581 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5641 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5701 tgcgcagcct gaatggcgaa tgcgatttat tcaacaaagc cgccgtcccg tcaagtcagc
     5761 gtaatgctct gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc
     5821 atcaaatgaa actgcaatat attcatatca ggattatcaa taccatattt ttgaaaaagc
     5881 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     5941 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     6001 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     6061 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     6121 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     6181 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     6241 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     6301 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     6361 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     6421 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     6481 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     6541 tacccatata aatcagcatc catgttggaa tttaatcgcg gcttcgagca agacgtttcc
     6601 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     6661 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     6721 tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc
     6781 gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac
     6841 aacaaagctc tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag
     6901 gcctggtatg agtcagcaac accttcttca cgaggcagac ctctcgacgg agttccactg
     6961 agcgtcagac
//
