LOCUS       Exported                6970 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human KMT2B.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_KMT2B
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 12, 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"
     primer_bind     complement(214..231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(385..973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(465..484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1342..1361)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1353..2168
                     /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     complement(1426..1445)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2372..2394
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2386..2403
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2387..2403
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3174..3968)
                     /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(3285..3304)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3895..3914
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3981..4037)
                     /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             complement(4053..4076)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4080..4103)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4107..4130)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5164..6294)
                     /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"
     primer_bind     complement(6370..6394)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6391..6593)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6420..6440)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(6770..6786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6770..6786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6783..6805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6794..6810
                     /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)."
     promoter        complement(6818..6848)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6863..6884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cggtttgcgt attgggaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca
       61 acgcagaccg ttccgtggca aagcaaaagt tcaaaatcag taaccgtcag tgccgataag
      121 ttcaaagtta aacctggtgt tgataccaac attgaaacgt tgatcgaaaa cgcgctgaaa
      181 aacgctgctg aatgtgcgag cttcttccgc ttcctcgctc actgactcgc tgcgctcggt
      241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
      841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaactc
     1021 cgtcgagagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc
     1081 catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc
     1141 agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg
     1201 tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc
     1261 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     1321 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     1381 gctcgaagcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1441 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     1501 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1561 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1621 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     1681 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1741 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     1801 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     1861 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     1921 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     1981 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2041 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2101 aatatggtat tgataatcct gatatgaata tattgcagtt tcatttgatg ctcgatgagt
     2161 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     2221 gggacggcgg ctttgttgaa taaatcgcat tcgccattca ggctgcgcaa ctgttgggaa
     2281 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     2341 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     2401 agtgaacgac ggccagtgaa ttggaggcta cagtcagtgg agaggacttt ccaagataaa
     2461 gaactttgct gactactaag cccagtatac aattagcact gaagtacttc ttgaagtaca
     2521 atcctaattg ggcaaagacc caacagatag cctcactgct cttcgcccta gaagggccta
     2581 ttccaacata cacacagggc cacatgtggg agggctgacc ccaggtgcct ctgggccacc
     2641 agaatcccct ctgggaaaca accctgctac agacagttgc tgtgttgtgt tgttctgaag
     2701 ccgtaacaga ccccatgact gacttctaat tctgaatgat ggcctcacct acctgttgag
     2761 gctgatgaac ttccttgggt catgatgggt gggtgtgtag aagtggggtt cctacctgcc
     2821 cccacagcct acaaatggga agaccacctc tccaaacatt gacagacttt attgtggggg
     2881 ggttcccacc tgggacccca cctcttaaat aaaaaagtgc atagaaaaga agggatttag
     2941 aaacctttgt acaatatcta catcctgggc ccccagggca gaaagggggc gattgctgga
     3001 tgggctggag gggtggaggc agggcccttg tcacctgcat gcccacatcc accctgaaca
     3061 tgagggtggg ggtgagatgc tctgggaggg gctagggagc ccccaggcta ggggcaagat
     3121 ggcagcgacg gcagcaggag gtgtgagggg tcgtggtggg caggatgttc gaatcagaag
     3181 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     3241 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     3301 aacgctatgt cctgatagcg gtcggccaca cccagccggc cacagtcgat gaatccagaa
     3361 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     3421 tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     3481 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     3541 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     3601 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     3661 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     3721 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     3781 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     3841 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     3901 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     3961 tcaatcatgc gaaacgatcc aggtccaggg ttctcctcca cgtctccagc ctgcttcagc
     4021 aggctgaagt tagtagctcc gcttcctgaa accttatcgt cgtcatcctt gtaatctccc
     4081 ttatcgtcgt catccttgta atcgccctta tcgtcgtcat ccttgtaatc tccagaacca
     4141 cctccaccgg aacctcctcc gctcccgtta aggaaccgac ggcagcgctt ggcgccacag
     4201 ttgcagggca gcttgttgct ggcatcctcg atggggaact tgtagtcgta ggtgagctcc
     4261 tcaccacgca ggatgcggcg cagggcgaag ataacaatgt gtttctggcc ctccacgtgg
     4321 atgacccgag agaagcagtt gggctcacag gagtggttga tgaagcgggc ggcattgcca
     4381 tgcatcgtgg cgtccactac atcaaagtca tccatgcgga acatatagca cccgatgccc
     4441 tgcagggatg aggtggggag atcaggtgct ctgtccccgc ggacaggctc gcttcaccca
     4501 ctgtcttccc acagccactg ggagcccacc ttcccatcgt agaacttctc ccgcttgtca
     4561 gtcaacaccg agcggatgac aatgccagag tactcgatga ccatctcccc cgcgtcgatg
     4621 ttgcgcttac agaacaggcc tcgcccgtgg atggctgatc tgcagggcag gatgaacagg
     4681 gaactcaggg gggaggggga cagaaaccag acagtacccc ctacccagtg cacctgtccg
     4741 tccacccaag gggcatcctc ccccccaacc ccactcacct gtagacaccc acagcttctt
     4801 tggacgtctt cttaaggtga cgaaaacgca tggccatggg cagctccagg ctggtggcac
     4861 gtctggtgag ggacagctgg gctgcaggtc ggggagagga gtgttagtaa ccgcgacgct
     4921 ggtgggggtc ggcttgttaa gaggggcgct gctaacgctg caagagtggg ttgtcagcgt
     4981 ggggccggtg ctactggaat cgataccggc atgattgaca gcctgggcga ggatgtcacc
     5041 tgatggtgat aagaatacac gggagactta gtacggtttc acaggcgtga cacgtttatt
     5101 gagtaggatt acagagtata acatagagta taatatagag tatacaatag tgacgtggga
     5161 tcctcagtta gcctccccca tctcccgggc aaacgtgcgc gccaggtcgc agatcgtcgg
     5221 tatggagccg ggggtggtga cgtgggtctg gaccatcccg gaggtaagtt gcagcagggc
     5281 gtcccggcag ccggcgggcg attggtcgta atccaggata aagacgtgca tgggacggag
     5341 gcgtttggcc aagacgtcca aggcccaggc aaacacgttg tacaggtcgc cgttgggggc
     5401 cagcaactcg ggggcccgaa acagggtaaa taacgtgtcc ccgatatggg gtcgtgggcc
     5461 cgcgttgctc tggggctcgg caccctgggg cggcacggcc gtccccgaaa gctgtcccca
     5521 atcctcccgc cacgacccgc cgccctgcag ataccgcacc gtattggcaa gcagcccgta
     5581 aacgcggcga atcgcggcca gcatagccag gtcaagccgc tcgccggggc gctggcgttt
     5641 ggccaggcgg tcgatgtgtc tgtcctccgg aagggccccc aacacgatgt ttgtgccggg
     5701 caaggtcggc gggatgaggg ccacgaacgc cagcacggcc tggggggtca tgctgcccat
     5761 aaggtatcgc gcggccgggt agcacaggag ggcggcgatg ggatggcggt cgaagatgag
     5821 ggtgagggcc gggggcgggg catgtgagct cccagcctcc cccccgatat gaggagccag
     5881 aacggcgtcg gtcacggcat aaggcatgcc cattgttatc tgggcgcttg tcattaccac
     5941 cgccgcgtcc ccggccgata tctcaccctg gtcgaggcgg tgttgtgtgg tgtagatgtt
     6001 cgcgattgtc tcggaagccc ccagcacctg ccagtaagtc atcggctcgg gtacgtagac
     6061 gatatcgtcg cgcgaaccca gggccaccag cagttgcgtg gtggtggttt tccccatccc
     6121 gtgaggaccg tctatataaa cccgcagtag cgtgggcatt ttctgctcca ggcggacttc
     6181 cgtggcttct tgctgccggc gagggcgcaa cgccgtacgt cggttgctat ggccgcgaga
     6241 acgcgcagcc tggtcgaacg cagacgcgtg ttgatggcag gggtacgaag ccatggtcgg
     6301 gatccccagg ctggatcggt cccggtgtct cctatggagg tcaaaacagc gtggatggcg
     6361 tctccaggcg atctgacggt tcactaaacg agctctgctt atatagacct cccaccgtac
     6421 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     6481 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttgaaatccc
     6541 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     6601 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     6661 tgggcataat gccagggaat tgacctgcag gcatgcaagc taattcgagc tcggtacccg
     6721 gggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt
     6781 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6841 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6901 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6961 cggggagagg
//
