LOCUS       sequence-260786-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human SSX2B.
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_SSX2B
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    18..39
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(54..59,60..77,78..84)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    92..108
                     /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     97..119
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     116..132
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     116..132
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        309..511
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     462..482
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     508..532
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             608..1738
                     /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             2772..2795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2799..2822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2826..2849
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2865..2921
                     /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             2934..3728
                     /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(2988..3007)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3598..3617
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4499..4515)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4508..4530)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4734..5549)
                     /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     5457..5476
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5541..5560
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5929..6517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6418..6437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6671..6688
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
       61 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
      121 aacagctatg accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc
      181 ccgggtaccg agctcgaatt agcttgcatg cctgcaggtc aattccctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      421 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      481 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
      541 acgccatcca cgctgttttg acctccatag gagacaccgg gaccgatcca gcctggggat
      601 cccgaccatg gcttcgtacc cctgccatca acacgcgtct gcgttcgacc aggctgcgcg
      661 ttctcgcggc catagcaacc gacgtacggc gttgcgccct cgccggcagc aagaagccac
      721 ggaagtccgc ctggagcaga aaatgcccac gctactgcgg gtttatatag acggtcctca
      781 cgggatgggg aaaaccacca ccacgcaact gctggtggcc ctgggttcgc gcgacgatat
      841 cgtctacgta cccgagccga tgacttactg gcaggtgctg ggggcttccg agacaatcgc
      901 gaacatctac accacacaac accgcctcga ccagggtgag atatcggccg gggacgcggc
      961 ggtggtaatg acaagcgccc agataacaat gggcatgcct tatgccgtga ccgacgccgt
     1021 tctggctcct catatcgggg gggaggctgg gagctcacat gccccgcccc cggccctcac
     1081 cctcatcttc gaccgccatc ccatcgccgc cctcctgtgc tacccggccg cgcgatacct
     1141 tatgggcagc atgacccccc aggccgtgct ggcgttcgtg gccctcatcc cgccgacctt
     1201 gcccggcaca aacatcgtgt tgggggccct tccggaggac agacacatcg accgcctggc
     1261 caaacgccag cgccccggcg agcggcttga cctggctatg ctggccgcga ttcgccgcgt
     1321 ttacgggctg cttgccaata cggtgcggta tctgcagggc ggcgggtcgt ggcgggagga
     1381 ttggggacag ctttcgggga cggccgtgcc gccccagggt gccgagcccc agagcaacgc
     1441 gggcccacga ccccatatcg gggacacgtt atttaccctg tttcgggccc ccgagttgct
     1501 ggcccccaac ggcgacctgt acaacgtgtt tgcctgggcc ttggacgtct tggccaaacg
     1561 cctccgtccc atgcacgtct ttatcctgga ttacgaccaa tcgcccgccg gctgccggga
     1621 cgccctgctg caacttacct ccgggatggt ccagacccac gtcaccaccc ccggctccat
     1681 accgacgatc tgcgacctgg cgcgcacgtt tgcccgggag atgggggagg ctaactgagg
     1741 atcccacgtc actattgtat actctatatt atactctatg ttatactctg taatcctact
     1801 caataaacgt gtcacgcctg tgaaaccgta ctaagtctcc cgtgtattct tatcaccatc
     1861 aggtgacatc ctcgcccagg ctgtcaatca tgccggtatc gattccagta gcaccggccc
     1921 cacgctgaca acccactctt gcagcgttag cagcgcccct cttaacaagc cgacccccac
     1981 cagcgtcgcg gttactaaca ctcctctccc cgacctgcag cccagcttga ggatggatac
     2041 cctattttcc atgatgtgat tattacccat tgcatgcctg tatcaaagca tctcatgtac
     2101 cccataagaa tatacacctg ctgcgtaccc acaaaataaa gaacaaaaat tattttaaaa
     2161 cactaaaaaa caataaaatc aattgcagta ataatatcaa agctcaccga tcacagatca
     2221 ctatagcaga tataataata atggaaaagt gtaaaattgg tgagaattaa aaaaaaaaat
     2281 tgcagcatct gcgaagcagt atgaacatga ggtgcaacaa aacaaggtac gcctgtgtgg
     2341 ccttaacaaa tacgtgctgg atgaaaggag gtatggggga atgttcccgt aagtgaagag
     2401 gttgggaatc taagcctgag aagggaagga gccagaagct aaaactttaa ttggcatttg
     2461 gcctatgttg gtgtggatct aaggtctcag cctctctaag ccagagaatg tgaaaaactg
     2521 gatgaagaag gcccatgggc acttgggagg aagcaggcat ctcctttttt tgagtaaaca
     2581 gagcctaaca ctctccaacc tacccaaccc tcactttcca actattctcc atcacaggac
     2641 ccaaaagggg ggaacatgcc tggacccaca gactgcgtga gagaaaacag ctggtgattt
     2701 atgaagagat cagcgaccct gaggaagatg acgaggggag cggaggaggt tccggtggag
     2761 gtggttctgg agattacaag gatgacgacg ataagggcga ttacaaggat gacgacgata
     2821 agggagatta caaggatgac gacgataagg tttcaggaag cggagctact aacttcagcc
     2881 tgctgaagca ggctggagac gtggaggaga accctggacc tggatcgttt cgcatgattg
     2941 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3001 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3061 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3121 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3181 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3241 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3301 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3361 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3421 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3481 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3541 tttctggatt catcgactgt ggccggctgg gtgtggccga ccgctatcag gacatagcgt
     3601 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3661 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3721 tcttctgatt cgaacatcac tcatccccca catccctgca gacgtgctat tctgttatga
     3781 tactggtatc ccatctgtca cttgctcccc aaatcattcc cttcttacaa ttttctactg
     3841 tacagcattg aggctgaacg atgagagatt tcccatgctc tttctactcc ctgccctgta
     3901 tatatccggg gatcctccct acccaggatg ctgtggggtc ccaaacccca agtaagccct
     3961 gatatgcggg ccacaccttt ctctagccta ggaattgata acccaggcga ggaagtcact
     4021 gtggcatgaa cagatggttc acttcgagga accgtggaag gcgtgtgcag gtcctgagat
     4081 agggcagaat cggagtgtgc agggtctgca ggtcaggagg agttgagatt gcgttgccac
     4141 gtggtgggaa ctcactgcca cttatttcct tctctcttct tgcctcagcc tcagggatac
     4201 gacacatgcc catgatgaga agcagaacgt ggtgaccttt cacgaacatg ggcatggctg
     4261 cggacccctc gtcatcaggt gcatagcaag tgaaagcaag tgttcacaac agtgaaaagt
     4321 tgagcgtcat ttttcttagt gtgccaagag ttcgatgtta gcgtttacgt tgtattttct
     4381 tacactgtgt cattctgtta gatactaaca ttttcattga tgagcaagac atacttaatg
     4441 catattcttg gaaagtcctc tccactgact gtagcctcca attcactggc cgtcgttcac
     4501 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     4561 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     4621 cttcccaaca gttgcgcagc ctgaatggcg aatgcgattt attcaacaaa gccgccgtcc
     4681 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
     4741 aactcatcga gcatcaaatg aaactgcaat atattcatat caggattatc aataccatat
     4801 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     4861 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     4921 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     4981 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     5041 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     5101 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     5161 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     5221 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     5281 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     5341 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     5401 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     5461 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcttcgag
     5521 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     5581 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     5641 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     5701 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     5761 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     5821 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctctcgac
     5881 ggagttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5941 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6001 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6061 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6121 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6181 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6241 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6301 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6361 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6421 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6481 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6541 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6601 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6661 gaccgagcgc agcgagtcag tgagcgagga agcggaagaa gctcgcacat tcagcagcgt
     6721 ttttcagcgc gttttcgatc aacgtttcaa tgttggtatc aacaccaggt ttaactttga
     6781 acttatcggc actgacggtt actgattttg aacttttgct ttgccacgga acggtctgcg
     6841 ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcccaa tacgcaaacc
     6901 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6961 gaaagcgggc
//
