LOCUS       sequence-168172-        6966 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor vector for 3' FLAG tag of human TEAD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  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 6966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome Res.
            2015 Sep 9."
COMMENT     Sequence Label: pFETCh_TEAD1
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             745..768
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             772..795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             799..822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             838..894
                     /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             907..1701
                     /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(961..980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1571..1590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2468..2484)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2477..2499)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2703..3518)
                     /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
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3426..3445
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3510..3529
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3898..4486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4387..4406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4640..4657
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4957..4978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4993..4998,4999..5016,5017..5023)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5031..5047
                     /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     5036..5058
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5055..5071
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5055..5071
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5248..5450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5401..5421
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5447..5471
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5547..6677
                     /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 acaagattta gcagaagttt aagactagga tgggaaaaag agtggatttt aaatgttctc
       61 atcacaaaga agtacgtgaa gtgcttggta tgttagcctg atgtaatcgt cccacaaggt
      121 atatacatct atcatcacat tgtagcccat aaatatatgt actgcaagac tttattcttt
      181 taggtaggga ctggaattat tacttgtcaa tttaaaaatt cagataaaaa ataagacaat
      241 ttcttagagt aaatgattgc cttgcgggac agaattaaag agaattcttt gttgtaaaga
      301 aatcagcccc ctaaaaaaca aaaacacaaa tgtgctaatt tttgtggtcc tttcttgggc
      361 atagattatg agtgacccta tctagaaggt atttcttact tatgataacc acgtttagat
      421 gggcttcagg tcatagttgt agtgttctga atctgtgtta gaggacacag agtgaggctg
      481 agcaaatctc tcttggactt aagacttgtt cttctccaat taagtcatag tcgtcataca
      541 cagatggaat atgttttcat ccttttggta ttatatactt tttttttatc ttaacaggtg
      601 gtaacaaaca gggatacaca agaaactcta ctctgcatgg cctgtgtgtt tgaagtttca
      661 aatagtgaac acggagcaca acatcatatt tacaggcttg taaaggacgg gagcggagga
      721 ggttccggtg gaggtggttc tggagattac aaggatgacg acgataaggg cgattacaag
      781 gatgacgacg ataagggaga ttacaaggat gacgacgata aggtttcagg aagcggagct
      841 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggatcg
      901 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
      961 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     1021 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     1081 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     1141 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     1201 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     1261 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     1321 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     1381 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     1441 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     1501 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc cgaccgctat
     1561 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     1621 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     1681 cttcttgacg agttcttctg attcgaacat cacatggtta tttatatata tagatatctg
     1741 tatatacaca cacacatatg tgcacacaca cactctctct ccattatcga acgactgact
     1801 gtaaacctca ccacacaggg tggtgccctg gccccgaggt caccccgact tttctaaatc
     1861 ttgtttgagt gaagtcattt tttcatgtgt tcatactatc attgtagctg tgaagttctg
     1921 gtacagttgt aaaaagagaa attgagttgt ttctctatgt tcttcagatg tgcagcccac
     1981 aattcctcgg gaaaggtgaa cctgaacaac ccaagtctct ctctgcagag ccctgtttct
     2041 aattgtggta gaaaatattg agacagagca tttgccatgg gacatttaca gcctttatac
     2101 aaatgtattt agttctcttt tttccaacat aaaattcttg ttttaagata caagtaaaat
     2161 taatctttaa atataaatgt aaattagtac acaaaactaa gaatctttag acttatcttt
     2221 gtaactaatt agggtggaag ttatgaaaga atgtaattca ctaaattatt ttttaaatga
     2281 aacctttttt tttctttttg aaaccaaatg ttaaactata gccttaagaa atgcttggta
     2341 gaagtgtcct aatgagacaa atttgtactt ttatcctcaa ggttaacact aatctcctaa
     2401 tccattaaac tcttgcttgg aaagtcctct ccactgactg tagcctccaa ttcactggcc
     2461 gtcgttcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     2521 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     2581 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgcgattta ttcaacaaag
     2641 ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct
     2701 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2761 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2821 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2881 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2941 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     3001 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     3061 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     3121 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     3181 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     3241 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     3301 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3361 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3421 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3481 ggcttcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     3541 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     3601 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     3661 gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa
     3721 atcaccaact ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg
     3781 ctggatgatg gggcgattca ggcctggtat gagtcagcaa caccttcttc acgaggcaga
     3841 cctctcgacg gagttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4561 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4621 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagaag ctcgcacatt
     4681 cagcagcgtt tttcagcgcg ttttcgatca acgtttcaat gttggtatca acaccaggtt
     4741 taactttgaa cttatcggca ctgacggtta ctgattttga acttttgctt tgccacggaa
     4801 cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcccaat
     4861 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4921 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     4981 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5041 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
     5101 ggtcgactct agaggatccc cgggtaccga gctcgaatta gcttgcatgc ctgcaggtca
     5161 attccctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     5221 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     5281 ggatagcggt ttgactcacg gggatttcaa gtctccaccc cattgacgtc aatgggagtt
     5341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     5401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     5461 accgtcagat cgcctggaga cgccatccac gctgttttga cctccatagg agacaccggg
     5521 accgatccag cctggggatc ccgaccatgg cttcgtaccc ctgccatcaa cacgcgtctg
     5581 cgttcgacca ggctgcgcgt tctcgcggcc atagcaaccg acgtacggcg ttgcgccctc
     5641 gccggcagca agaagccacg gaagtccgcc tggagcagaa aatgcccacg ctactgcggg
     5701 tttatataga cggtcctcac gggatgggga aaaccaccac cacgcaactg ctggtggccc
     5761 tgggttcgcg cgacgatatc gtctacgtac ccgagccgat gacttactgg caggtgctgg
     5821 gggcttccga gacaatcgcg aacatctaca ccacacaaca ccgcctcgac cagggtgaga
     5881 tatcggccgg ggacgcggcg gtggtaatga caagcgccca gataacaatg ggcatgcctt
     5941 atgccgtgac cgacgccgtt ctggctcctc atatcggggg ggaggctggg agctcacatg
     6001 ccccgccccc ggccctcacc ctcatcttcg accgccatcc catcgccgcc ctcctgtgct
     6061 acccggccgc gcgatacctt atgggcagca tgacccccca ggccgtgctg gcgttcgtgg
     6121 ccctcatccc gccgaccttg cccggcacaa acatcgtgtt gggggccctt ccggaggaca
     6181 gacacatcga ccgcctggcc aaacgccagc gccccggcga gcggcttgac ctggctatgc
     6241 tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac ggtgcggtat ctgcagggcg
     6301 gcgggtcgtg gcgggaggat tggggacagc tttcggggac ggccgtgccg ccccagggtg
     6361 ccgagcccca gagcaacgcg ggcccacgac cccatatcgg ggacacgtta tttaccctgt
     6421 ttcgggcccc cgagttgctg gcccccaacg gcgacctgta caacgtgttt gcctgggcct
     6481 tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt tatcctggat tacgaccaat
     6541 cgcccgccgg ctgccgggac gccctgctgc aacttacctc cgggatggtc cagacccacg
     6601 tcaccacccc cggctccata ccgacgatct gcgacctggc gcgcacgttt gcccgggaga
     6661 tgggggaggc taactgagga tcccacgtca ctattgtata ctctatatta tactctatgt
     6721 tatactctgt aatcctactc aataaacgtg tcacgcctgt gaaaccgtac taagtctccc
     6781 gtgtattctt atcaccatca ggtgacatcc tcgcccaggc tgtcaatcat gccggtatcg
     6841 attccagtag caccggcccc acgctgacaa cccactcttg cagcgttagc agcgcccctc
     6901 ttaacaagcc gacccccacc agcgtcgcgg ttactaacac tcctctcccc gacctgcagc
     6961 ccagct
//
