LOCUS       sequence-199544-        6889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human NFIA_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6889)
  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 6889)
  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_NFIA_iso1
FEATURES             Location/Qualifiers
     source          1..6889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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     2391..2407
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3097..3891)
                     /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(3208..3227)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3818..3837
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3904..3960)
                     /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(3976..3999)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4003..4026)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4030..4053)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5087..6217)
                     /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(6293..6317)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6314..6516)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6343..6363)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(6693..6709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6693..6709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6706..6728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6717..6733
                     /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(join(6741..6747,6748..6765,6766..6771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6786..6807
                     /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 gctacagtca gtggagagga ctttccaaga
     2461 ttgtccttgc gggccatgtt atatacttaa ctgctgactg ctgaaccaaa acaaactaaa
     2521 ggaacaaata aaaaaggaat ataactgcat tttctttttc caatatgtta cagaaaaaca
     2581 ttgccctttg actgtcccgt tatatattgc agatctcagc tactacaaaa gatttagttc
     2641 tccatgtgcc tcctcaatgc tcaaggagga tcacacctgt gtgggtaaat tcatgaaata
     2701 ctgtgaaaca gccaggctcc gtaaggaaag tgtgttacaa caggaagtca gtgagcaagg
     2761 gtagtcctcc gctagccacg tcacctaatg ctcacgctac catggaaaca cttcaaatgt
     2821 tacaaagccc aaccatttcc agcaccatat gagaaattac aacagtccct aaagtatttt
     2881 aaaaaaaaaa aaatcccaaa aggttatggc ctttgctgtt tgaccataat gcttgctgtt
     2941 tccatgtaca agttgatttg aatcggtttt tctccccttc cgctgatagt gaagttgtaa
     3001 ctgcgctggg ttgaggttgc gtccatgtta cagagttgag aaggggtcag gtggtctgtc
     3061 tggtggatgg tgggacgctg caacttgatg ttcgaatcag aagaactcgt caagaaggcg
     3121 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3181 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3241 gcggtcggcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3301 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     3361 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3421 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3481 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3541 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     3601 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     3661 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     3721 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     3781 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     3841 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     3901 tccaggtcca gggttctcct ccacgtctcc agcctgcttc agcaggctga agttagtagc
     3961 tccgcttcct gaaaccttat cgtcgtcatc cttgtaatct cccttatcgt cgtcatcctt
     4021 gtaatcgccc ttatcgtcgt catccttgta atctccagaa ccacctccac cggaacctcc
     4081 tccgctcccg cctagatacc aggactggaa gacaaaagga aaaatacaat taaaatcaca
     4141 atttgtaaaa acgtgtgttt tagaagttgc cttttcaacc acgaaatcag gatctacgtt
     4201 agggatgcgg gagggattcg ctgggaaaag taaaggcacg acagacaata aaaaatcctg
     4261 tgacactgaa actgatctta ttcttctgaa attataattc attgtcttca ttggggtttt
     4321 ccaacattct gagtatatct tacagaatga ggtatttctg ttcacttgaa atacaaataa
     4381 gggttttgtt aactgatggg ggcagttata ggagggcagg aaacttgacc atcttgttta
     4441 tcattcaatc ctcagtgcct agcccagtgc ctggaataca gaacgttctt aatgaagaag
     4501 agttgaatga atggagaatg ggtaaaggga ggggtttcat gtggcaatga atggctactc
     4561 tgatgatttt caacatctcc tttcaaatga ggtgcaggca ctgaagccat gattgtgaca
     4621 ctctcctaac aggcccagat ggtcaatagc gacacaagaa taagtggagg taaggtggaa
     4681 aaagtgctgg acccagtctt cctgggtgat tttgagcaag tcacagtatt tttccttcct
     4741 tacatatgta ggattttatg agggaagatg ctcctttcag cccaatatat ctaaaatagc
     4801 tgggctgcag gtcggggaga ggagtgttag taaccgcgac gctggtgggg gtcggcttgt
     4861 taagaggggc gctgctaacg ctgcaagagt gggttgtcag cgtggggccg gtgctactgg
     4921 aatcgatacc ggcatgattg acagcctggg cgaggatgtc acctgatggt gataagaata
     4981 cacgggagac ttagtacggt ttcacaggcg tgacacgttt attgagtagg attacagagt
     5041 ataacataga gtataatata gagtatacaa tagtgacgtg ggatcctcag ttagcctccc
     5101 ccatctcccg ggcaaacgtg cgcgccaggt cgcagatcgt cggtatggag ccgggggtgg
     5161 tgacgtgggt ctggaccatc ccggaggtaa gttgcagcag ggcgtcccgg cagccggcgg
     5221 gcgattggtc gtaatccagg ataaagacgt gcatgggacg gaggcgtttg gccaagacgt
     5281 ccaaggccca ggcaaacacg ttgtacaggt cgccgttggg ggccagcaac tcgggggccc
     5341 gaaacagggt aaataacgtg tccccgatat ggggtcgtgg gcccgcgttg ctctggggct
     5401 cggcaccctg gggcggcacg gccgtccccg aaagctgtcc ccaatcctcc cgccacgacc
     5461 cgccgccctg cagataccgc accgtattgg caagcagccc gtaaacgcgg cgaatcgcgg
     5521 ccagcatagc caggtcaagc cgctcgccgg ggcgctggcg tttggccagg cggtcgatgt
     5581 gtctgtcctc cggaagggcc cccaacacga tgtttgtgcc gggcaaggtc ggcgggatga
     5641 gggccacgaa cgccagcacg gcctgggggg tcatgctgcc cataaggtat cgcgcggccg
     5701 ggtagcacag gagggcggcg atgggatggc ggtcgaagat gagggtgagg gccgggggcg
     5761 gggcatgtga gctcccagcc tcccccccga tatgaggagc cagaacggcg tcggtcacgg
     5821 cataaggcat gcccattgtt atctgggcgc ttgtcattac caccgccgcg tccccggccg
     5881 atatctcacc ctggtcgagg cggtgttgtg tggtgtagat gttcgcgatt gtctcggaag
     5941 cccccagcac ctgccagtaa gtcatcggct cgggtacgta gacgatatcg tcgcgcgaac
     6001 ccagggccac cagcagttgc gtggtggtgg ttttccccat cccgtgagga ccgtctatat
     6061 aaacccgcag tagcgtgggc attttctgct ccaggcggac ttccgtggct tcttgctgcc
     6121 ggcgagggcg caacgccgta cgtcggttgc tatggccgcg agaacgcgca gcctggtcga
     6181 acgcagacgc gtgttgatgg caggggtacg aagccatggt cgggatcccc aggctggatc
     6241 ggtcccggtg tctcctatgg aggtcaaaac agcgtggatg gcgtctccag gcgatctgac
     6301 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     6361 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     6421 acaaactccc attgacgtca atggggtgga gacttgaaat ccccgtgagt caaaccgcta
     6481 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     6541 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggg
     6601 aattgacctg caggcatgca agctaattcg agctcggtac ccggggatcc tctagagtcg
     6661 acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6721 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6781 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6841 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcgggga
//
