LOCUS       Exported                6968 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human ZNF652.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_ZNF652
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6968)
  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 6968)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6968
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             747..770
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             774..797
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             801..824
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             840..896
                     /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             909..1703
                     /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(963..982)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1573..1592
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2473..2490)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2473..2489)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2482..2504)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2708..3523)
                     /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     3431..3450
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3515..3534
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3903..4491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4392..4411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4645..4662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4962..4983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4998..5028
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5036..5052
                     /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     5041..5063
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5060..5076
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5060..5076
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5253..5455
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5406..5426
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5452..5476
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5552..6682
                     /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"
ORIGIN
        1 gctgtctcat aggccaggcg cagtggctca cacctgtaat cgcagcactt tgggaggcca
       61 agatgggagg atcacttgag gccaggagtt cgagaccagc ctgatcaaca tagtgagacc
      121 ccatttctat taaaaaaaaa aagcttgagc acacaattac cacctaatat gttaatcatt
      181 tttatgtctg tgttcatctc aggagagaaa ccctttatct gtgaaatctg tggcaaaagc
      241 ttcaccagcc gccccaacat gaagagacac cgcagaactc acacaggcga gaagccctat
      301 ccatgtgatg tgtgtggcca gcggttccgc ttctcgaaca tgcttaaggc ccacaaggag
      361 aagtgctttc gggtgaccag ccccgtgaat gtgccacctg ctgtccagat cccacttaca
      421 acttccccag ccaccccagt tccttctgtg gtgaacacag ccacaacccc aaccccccaa
      481 tcaatatgaa tcctgtaagc actcttcccc ctcggcccat cccccacccc ttctcacacc
      541 tgcacatcca cccacaccct caccacccac accaccttcc catccctcca gtccctcacc
      601 tcccgccacc tccagctctc tttaagagtg agcctttaaa tcatagaggc cagagtgagg
      661 acaactttct gcggcacctg gcagagaaga acagttcagc acagcatcat gggagcggag
      721 gaggttccgg tggaggtggt tctggagatt acaaggatga cgacgataag ggcgattaca
      781 aggatgacga cgataaggga gattacaagg atgacgacga taaggtttca ggaagcggag
      841 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctggat
      901 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      961 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     1021 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     1081 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     1141 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     1201 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     1261 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     1321 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     1381 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     1441 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     1501 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gccgaccgct
     1561 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     1621 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     1681 gccttcttga cgagttcttc tgattcgaac atctgtgttc tccaggagac acgtccccat
     1741 gtgtgagcgt gcacagaggg agtcggtgag catttcctta gttctcagac tctcctccgc
     1801 ctctaccaag agcttcgcca gtgtcactga atcaacagat ccaagggaat gaacaagaag
     1861 actaccttga gctcacagag ggactgtcat ctaaaccagg ggaaccactg aactaaagcc
     1921 caatttgctt gcattttctg atgactttta taagtttcaa atactcagac ttgtggaatt
     1981 ttataatttc atatcagctg atgaggtatg cttgctttta tatcccagac ttctcctcaa
     2041 agaggggata ggcctggttt cctttgtact aatcgggaag gctgtgagat taatccagag
     2101 cattaattgt atcactgtgt atggtaagga attcttttca gcttttggtg ccagctacag
     2161 agttgagctg gccatgtttc acagtatatc aagcattata gagaaagaca gaaattttct
     2221 tctttgtgta gctctcctaa cgcactcttt attttactac agaaattatt ttagagtttt
     2281 tgtatagact tctttggtag tctgtttcta aaatgaaatg tatttcaata agcagtgtaa
     2341 ttttttcagt gtagctgaac ctatgttgta aaatagaaaa aaatttttat aatcatcaaa
     2401 atgtgattct taaagatgca cttggaaagt cctctccact gactgtagcc tccaattcac
     2461 tggccgtcgt tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2521 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2581 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgcg atttattcaa
     2641 caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca
     2701 attctgatta gaaaaactca tcgagcatca aatgaaactg caatatattc atatcaggat
     2761 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     2821 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     2881 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     2941 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa
     3001 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     3061 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     3121 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     3181 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     3241 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     3301 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     3361 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     3421 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     3481 atcgcggctt cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     3541 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     3601 aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt
     3661 tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt
     3721 tcaaaatcac caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt
     3781 tctggctgga tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag
     3841 gcagacctct cgacggagtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3901 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3961 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4021 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4081 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4141 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4201 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4261 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4321 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4381 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4441 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4501 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4561 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4621 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agaagctcgc
     4681 acattcagca gcgtttttca gcgcgttttc gatcaacgtt tcaatgttgg tatcaacacc
     4741 aggtttaact ttgaacttat cggcactgac ggttactgat tttgaacttt tgctttgcca
     4801 cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc
     4861 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     4921 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4981 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     5041 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc
     5101 ctgcaggtcg actctagagg atccccgggt accgagctcg aattagcttg catgcctgca
     5161 ggtcaattcc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     5221 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     5281 ggcgtggata gcggtttgac tcacggggat ttcaagtctc caccccattg acgtcaatgg
     5341 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5401 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     5461 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc ataggagaca
     5521 ccgggaccga tccagcctgg ggatcccgac catggcttcg tacccctgcc atcaacacgc
     5581 gtctgcgttc gaccaggctg cgcgttctcg cggccatagc aaccgacgta cggcgttgcg
     5641 ccctcgccgg cagcaagaag ccacggaagt ccgcctggag cagaaaatgc ccacgctact
     5701 gcgggtttat atagacggtc ctcacgggat ggggaaaacc accaccacgc aactgctggt
     5761 ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag ccgatgactt actggcaggt
     5821 gctgggggct tccgagacaa tcgcgaacat ctacaccaca caacaccgcc tcgaccaggg
     5881 tgagatatcg gccggggacg cggcggtggt aatgacaagc gcccagataa caatgggcat
     5941 gccttatgcc gtgaccgacg ccgttctggc tcctcatatc gggggggagg ctgggagctc
     6001 acatgccccg cccccggccc tcaccctcat cttcgaccgc catcccatcg ccgccctcct
     6061 gtgctacccg gccgcgcgat accttatggg cagcatgacc ccccaggccg tgctggcgtt
     6121 cgtggccctc atcccgccga ccttgcccgg cacaaacatc gtgttggggg cccttccgga
     6181 ggacagacac atcgaccgcc tggccaaacg ccagcgcccc ggcgagcggc ttgacctggc
     6241 tatgctggcc gcgattcgcc gcgtttacgg gctgcttgcc aatacggtgc ggtatctgca
     6301 gggcggcggg tcgtggcggg aggattgggg acagctttcg gggacggccg tgccgcccca
     6361 gggtgccgag ccccagagca acgcgggccc acgaccccat atcggggaca cgttatttac
     6421 cctgtttcgg gcccccgagt tgctggcccc caacggcgac ctgtacaacg tgtttgcctg
     6481 ggccttggac gtcttggcca aacgcctccg tcccatgcac gtctttatcc tggattacga
     6541 ccaatcgccc gccggctgcc gggacgccct gctgcaactt acctccggga tggtccagac
     6601 ccacgtcacc acccccggct ccataccgac gatctgcgac ctggcgcgca cgtttgcccg
     6661 ggagatgggg gaggctaact gaggatccca cgtcactatt gtatactcta tattatactc
     6721 tatgttatac tctgtaatcc tactcaataa acgtgtcacg cctgtgaaac cgtactaagt
     6781 ctcccgtgta ttcttatcac catcaggtga catcctcgcc caggctgtca atcatgccgg
     6841 tatcgattcc agtagcaccg gccccacgct gacaacccac tcttgcagcg ttagcagcgc
     6901 ccctcttaac aagccgaccc ccaccagcgt cgcggttact aacactcctc tccccgacct
     6961 gcagccca
//
