LOCUS       Exported                6955 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human ARID4B.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_ARID4B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  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 6955)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6955
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(218..235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(389..977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(469..488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1346..1365)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1357..2172
                     /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(1430..1449)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2376..2398
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2390..2407
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2391..2407
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3163..3957)
                     /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(3274..3293)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3884..3903
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3970..4026)
                     /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(4042..4065)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4069..4092)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4096..4119)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5153..6283)
                     /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(6359..6383)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6380..6582)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6409..6429)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(6759..6775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6759..6775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6772..6794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6783..6799
                     /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(6807..6837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6852..6873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg aacttttgct gagttgaagg atcagatcac gcatcttccc
       61 gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcagtaaccg tcagtgccga
      121 taagttcaaa gttaaacctg gtgttgatac caacattgaa acgttgatcg aaaacgcgct
      181 gaaaaacgct gctgaatgtg cgagcttctt ccgcttcctc gctcactgac tcgctgcgct
      241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1021 actccgtcga gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc
     1081 gccccatcat ccagccagaa agtgagggag ccacggttga tgagagcttt gttgtaggtg
     1141 gaccagttgg tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga
     1201 tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag ccacgttgtg
     1261 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1321 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1381 tcttgctcga agccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1441 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     1501 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1561 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1621 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     1681 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1741 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1801 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1861 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc
     1921 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1981 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2041 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2101 caaaaatatg gtattgataa tcctgatatg aatatattgc agtttcattt gatgctcgat
     2161 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2221 tgacgggacg gcggctttgt tgaataaatc gcattcgcca ttcaggctgc gcaactgttg
     2281 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     2341 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     2401 ggccagtgaa cgacggccag tgaattggag gctacagcca gtggagagga ctttccaaga
     2461 aaaatactga tttatagaaa aatggcaaag tacagtagtt ccattccaat ttgaagggcc
     2521 atgaaaagcc actgcaagac cttttagcct aattcaaacc tgtaaacatg ttcagtcttt
     2581 tttaaagaga atctttaata tttggttacc aggattgatg tctaatatcc tgttaactgc
     2641 aggttttgaa tttattacat gtgcttgact tatacaatta aagccaccct aaggtgactt
     2701 gctttaaaaa acaattacct tgtacaaatg aaaataggtt catatttttt cataaataaa
     2761 tggaaaaata tcaaatgttc cagctttaac aaaatacaag ggaatacata tacagtaagt
     2821 tatcttaaca tgttctgacc caccccttaa ctatgcttac tgtattgtct ctacaggcag
     2881 tgaaaagcct ccatttgcca cagtggaagg ccttccagtt accagacaca caattcccag
     2941 acaagttgga aacaattatt gcttgaggaa aagcagttca ttgtactttt tcttcataaa
     3001 aaagtgcact taagtgccaa gtcagcttgt caagaaacaa tttttaaata taaaatagtg
     3061 cttgtctgat atttttttgt gccactgtgc agtataaaaa aaaagtggcc ctcaacagcc
     3121 attaagtgca aagtgcttta gcaagtcctg ctgatgttcg aatcagaaga actcgtcaag
     3181 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     3241 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     3301 ctgatagcgg tcggccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     3361 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     3421 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     3481 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     3541 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     3601 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     3661 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     3721 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     3781 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     3841 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     3901 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     3961 aaacgatcca ggtccagggt tctcctccac gtctccagcc tgcttcagca ggctgaagtt
     4021 agtagctccg cttcctgaaa ccttatcgtc gtcatccttg taatctccct tatcgtcgtc
     4081 atccttgtaa tcgcccttat cgtcgtcatc cttgtaatct ccagaaccac ctccaccgga
     4141 acctcctccg ctccccctgc actcaactga cattccattt tgtgatgcgc tggacattga
     4201 cggttcagct aaagttaaca taacagcagc tgttatggaa ctggatgaag gtgaagagga
     4261 ggatgaggat gtagcagcac ctaaggaaaa gggagaccaa accaagagct taataaaaat
     4321 ttatgtctaa caatagacag aaaatactag tcctctgaac taaaattccg ctatattgtc
     4381 caaaatcatt tagcttacaa caacagtggt caattctcac agttctacga tgtgaagaaa
     4441 ggaaaagcta gtgtagctca agaagctaga cagtgcccaa ctacagaagt aatgaaaagg
     4501 acagaccagg gaaggctcct gcgtgggtca aagagggaaa gaactgtgcc tagtctacaa
     4561 gacacagaaa taaaatacag aatacacttt ttgtaacagt gaaacaagta tcaaaacagg
     4621 aaaaatctgc aatccatcca aaaatcattt gtcccctcct aatccctcca tctttctgta
     4681 cttaggagtt ttcttgtaca tgtaaatggt gacaagagtg ggaggcttaa ttgcttcttc
     4741 aagttggatc aaaacagtgc atagctcaac tgttgcattc tccgccacta ggtttacaaa
     4801 gcaaataaca cattagttat tttcatcaga ggaagtctct cactgtttcc ttttttcgtt
     4861 tttagctggg ctgcaggtcg gggagaggag tgttagtaac cgcgacgctg gtgggggtcg
     4921 gcttgttaag aggggcgctg ctaacgctgc aagagtgggt tgtcagcgtg gggccggtgc
     4981 tactggaatc gataccggca tgattgacag cctgggcgag gatgtcacct gatggtgata
     5041 agaatacacg ggagacttag tacggtttca caggcgtgac acgtttattg agtaggatta
     5101 cagagtataa catagagtat aatatagagt atacaatagt gacgtgggat cctcagttag
     5161 cctcccccat ctcccgggca aacgtgcgcg ccaggtcgca gatcgtcggt atggagccgg
     5221 gggtggtgac gtgggtctgg accatcccgg aggtaagttg cagcagggcg tcccggcagc
     5281 cggcgggcga ttggtcgtaa tccaggataa agacgtgcat gggacggagg cgtttggcca
     5341 agacgtccaa ggcccaggca aacacgttgt acaggtcgcc gttgggggcc agcaactcgg
     5401 gggcccgaaa cagggtaaat aacgtgtccc cgatatgggg tcgtgggccc gcgttgctct
     5461 ggggctcggc accctggggc ggcacggccg tccccgaaag ctgtccccaa tcctcccgcc
     5521 acgacccgcc gccctgcaga taccgcaccg tattggcaag cagcccgtaa acgcggcgaa
     5581 tcgcggccag catagccagg tcaagccgct cgccggggcg ctggcgtttg gccaggcggt
     5641 cgatgtgtct gtcctccgga agggccccca acacgatgtt tgtgccgggc aaggtcggcg
     5701 ggatgagggc cacgaacgcc agcacggcct ggggggtcat gctgcccata aggtatcgcg
     5761 cggccgggta gcacaggagg gcggcgatgg gatggcggtc gaagatgagg gtgagggccg
     5821 ggggcggggc atgtgagctc ccagcctccc ccccgatatg aggagccaga acggcgtcgg
     5881 tcacggcata aggcatgccc attgttatct gggcgcttgt cattaccacc gccgcgtccc
     5941 cggccgatat ctcaccctgg tcgaggcggt gttgtgtggt gtagatgttc gcgattgtct
     6001 cggaagcccc cagcacctgc cagtaagtca tcggctcggg tacgtagacg atatcgtcgc
     6061 gcgaacccag ggccaccagc agttgcgtgg tggtggtttt ccccatcccg tgaggaccgt
     6121 ctatataaac ccgcagtagc gtgggcattt tctgctccag gcggacttcc gtggcttctt
     6181 gctgccggcg agggcgcaac gccgtacgtc ggttgctatg gccgcgagaa cgcgcagcct
     6241 ggtcgaacgc agacgcgtgt tgatggcagg ggtacgaagc catggtcggg atccccaggc
     6301 tggatcggtc ccggtgtctc ctatggaggt caaaacagcg tggatggcgt ctccaggcga
     6361 tctgacggtt cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     6421 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     6481 gccaaaacaa actcccattg acgtcaatgg ggtggagact tgaaatcccc gtgagtcaaa
     6541 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     6601 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     6661 ccagggaatt gacctgcagg catgcaagct aattcgagct cggtacccgg ggatcctcta
     6721 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     6781 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6841 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6901 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc gggga
//
