LOCUS       sequence-260784-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human SIX4_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 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_SIX4_iso1
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        379..581
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     532..552
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     578..602
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             678..1808
                     /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             2842..2865
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2869..2892
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2896..2919
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2935..2991
                     /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             3004..3798
                     /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(3058..3077)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3668..3687
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4569..4585)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4578..4600)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4804..5619)
                     /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     5527..5546
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5611..5630
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5999..6587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6488..6507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6741..6758
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc aggtcgactc
      241 tagaggatcc ccgggtaccg agctcgaatt agcttgcatg cctgcaggtc aattccctgg
      301 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      361 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      421 tttgactcac ggggatttca agtctccacc ccattgacgt caatgggagt ttgttttggc
      481 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      541 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      601 tcgcctggag acgccatcca cgctgttttg acctccatag gagacaccgg gaccgatcca
      661 gcctggggat cccgaccatg gcttcgtacc cctgccatca acacgcgtct gcgttcgacc
      721 aggctgcgcg ttctcgcggc catagcaacc gacgtacggc gttgcgccct cgccggcagc
      781 aagaagccac ggaagtccgc ctggagcaga aaatgcccac gctactgcgg gtttatatag
      841 acggtcctca cgggatgggg aaaaccacca ccacgcaact gctggtggcc ctgggttcgc
      901 gcgacgatat cgtctacgta cccgagccga tgacttactg gcaggtgctg ggggcttccg
      961 agacaatcgc gaacatctac accacacaac accgcctcga ccagggtgag atatcggccg
     1021 gggacgcggc ggtggtaatg acaagcgccc agataacaat gggcatgcct tatgccgtga
     1081 ccgacgccgt tctggctcct catatcgggg gggaggctgg gagctcacat gccccgcccc
     1141 cggccctcac cctcatcttc gaccgccatc ccatcgccgc cctcctgtgc tacccggccg
     1201 cgcgatacct tatgggcagc atgacccccc aggccgtgct ggcgttcgtg gccctcatcc
     1261 cgccgacctt gcccggcaca aacatcgtgt tgggggccct tccggaggac agacacatcg
     1321 accgcctggc caaacgccag cgccccggcg agcggcttga cctggctatg ctggccgcga
     1381 ttcgccgcgt ttacgggctg cttgccaata cggtgcggta tctgcagggc ggcgggtcgt
     1441 ggcgggagga ttggggacag ctttcgggga cggccgtgcc gccccagggt gccgagcccc
     1501 agagcaacgc gggcccacga ccccatatcg gggacacgtt atttaccctg tttcgggccc
     1561 ccgagttgct ggcccccaac ggcgacctgt acaacgtgtt tgcctgggcc ttggacgtct
     1621 tggccaaacg cctccgtccc atgcacgtct ttatcctgga ttacgaccaa tcgcccgccg
     1681 gctgccggga cgccctgctg caacttacct ccgggatggt ccagacccac gtcaccaccc
     1741 ccggctccat accgacgatc tgcgacctgg cgcgcacgtt tgcccgggag atgggggagg
     1801 ctaactgagg atcccacgtc actattgtat actctatatt atactctatg ttatactctg
     1861 taatcctact caataaacgt gtcacgcctg tgaaaccgta ctaagtctcc cgtgtattct
     1921 tatcaccatc aggtgacatc ctcgcccagg ctgtcaatca tgccggtatc gattccagta
     1981 gcaccggccc cacgctgaca acccactctt gcagcgttag cagcgcccct cttaacaagc
     2041 cgacccccac cagcgtcgcg gttactaaca ctcctctccc cgacctgcag cccagctagc
     2101 tctcttgctc ccagtgcagt ggtatacacg gttcctaata caggccagac tataggatct
     2161 gtgaaacagg aaggcttgga aaggagcctg gtattttctc agttgatgcc tgtcaatcag
     2221 aatgcacaag taaatgcaaa cctgtcttct gaaaacatct cggggagtgg cctgcatcca
     2281 ctggcctcct cattagttaa tgtatctcca actcacaatt tttctctcag tccctctaca
     2341 ctactaaatc ccactgagct aaaccgcgac attgccgata gccaaccaat gtctgcaccg
     2401 gtggcaagca aatctactgt gacatctgtc agcaacacta actatgcaac tcttcagaac
     2461 tgctccctta ttactggtca agacctattg tcagtcccta tgactcaggc tgcccttggg
     2521 gaaatagttc ctacagctga agatcaggta ggtcacccct ccccagcagt acatcaggat
     2581 tttgtccaag aacatcgttt ggttctgcaa tcggtagcta acatgaaaga gaatttctta
     2641 tcaaattctg agagcaaagc aacaagtagc ttaatgatgc tggactctaa atccaagtat
     2701 gtcttagatg gcatggttga tactgtctgt gaagacctgg aaacagacaa aaaagagctt
     2761 gccaagctcc agactgtcca gctggatgaa gatatgcaag acttagggag cggaggaggt
     2821 tccggtggag gtggttctgg agattacaag gatgacgacg ataagggcga ttacaaggat
     2881 gacgacgata agggagatta caaggatgac gacgataagg tttcaggaag cggagctact
     2941 aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc tggatcgttt
     3001 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3061 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3121 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3181 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3241 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3301 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3361 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3421 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3481 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3541 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3601 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggccga ccgctatcag
     3661 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3721 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3781 cttgacgagt tcttctgatt cgaacatcct ctttttggca tcagcggcaa atcttttcat
     3841 gaagccccaa ggacacaaaa cattttccca tttaaaggaa aacactctag ttttgcaagt
     3901 atatgcatac aagagacttt agattgatct gcatgaagat cacagttaag tatacaggag
     3961 tagaactgca ttattgcagc ctttttgttc acttataaat ttctctttta aatagatgga
     4021 gacaaaggac aaggtgaaat gtatcaagtc aaagtgaatc atttagttga ctctataatt
     4081 ctaaggtcaa aatggaactt gatagttttt taaattaaaa aatgtataca cctaacatag
     4141 aaaattaaag atagctgcag accattagaa ataatacaat tgtttttgtt tacttttact
     4201 ccatgggcat tgaaaaggtt aagaaacata aatggtacat atttttaaag ttaagtagca
     4261 tgcatatata tatgcacaca cacctctttt tcagcatttt ttgagaaagt cttgggtctc
     4321 aaacacattt gtctcaacac atttccaaat gtgatttcta atagctcagt gtggctgaga
     4381 aagtgcggaa ctccactaaa aactctgaaa cttcagtata aatgtaaata tatcagttct
     4441 ataagcagga tttgagtaat tacagtgaat tttattttta atgtatatgt ttctcttcag
     4501 gcgctatgaa ctgaaacttg gaaagtcctc tccactgact gtagcctcca attcactggc
     4561 cgtcgttcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4621 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4681 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgcgattt attcaacaaa
     4741 gccgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
     4801 tgattagaaa aactcatcga gcatcaaatg aaactgcaat atattcatat caggattatc
     4861 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     4921 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     4981 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     5041 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     5101 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     5161 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     5221 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     5281 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     5341 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     5401 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     5461 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     5521 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     5581 cggcttcgag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     5641 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     5701 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     5761 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     5821 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
     5881 gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag
     5941 acctctcgac ggagttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6121 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6661 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6721 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagaa gctcgcacat
     6781 tcagcagcgt ttttcagcgc gttttcgatc aacgtttcaa tgttggtatc aacaccaggt
     6841 ttaactttga acttatcggc actgacggtt actgattttg aacttttgct ttgccacgga
     6901 acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcccaa
     6961 tacgcaaacc
//
