LOCUS       sequence-171032-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human NFIL3.
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_NFIL3
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      46..634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     535..554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     788..805
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1105..1126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1141..1146,1147..1164,1165..1171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1179..1195
                     /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     1184..1206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1203..1219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1203..1219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1396..1598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1549..1569
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1595..1619
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1695..2825
                     /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             3859..3882
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3886..3909
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3913..3936
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3952..4008
                     /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             4021..4815
                     /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(4075..4094)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4685..4704
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5586..5602)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5595..5617)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5821..6636)
                     /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     6544..6563
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6628..6647
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
       61 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      121 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      181 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      241 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      301 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      361 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      421 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      481 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      541 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      601 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      661 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      721 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      781 cgagcgcagc gagtcagtga gcgaggaagc ggaagaagct cgcacattca gcagcgtttt
      841 tcagcgcgtt ttcgatcaac gtttcaatgt tggtatcaac accaggttta actttgaact
      901 tatcggcact gacggttact gattttgaac ttttgctttg ccacggaacg gtctgcgttg
      961 tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcccaatac gcaaaccgcc
     1021 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     1081 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     1141 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     1201 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag
     1261 aggatccccg ggtaccgagc tcgaattagc ttgcatgcct gcaggtcaat tccctggcat
     1321 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     1381 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     1441 gactcacggg gatttcaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     1501 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     1561 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     1621 cctggagacg ccatccacgc tgttttgacc tccataggag acaccgggac cgatccagcc
     1681 tggggatccc gaccatggct tcgtacccct gccatcaaca cgcgtctgcg ttcgaccagg
     1741 ctgcgcgttc tcgcggccat agcaaccgac gtacggcgtt gcgccctcgc cggcagcaag
     1801 aagccacgga agtccgcctg gagcagaaaa tgcccacgct actgcgggtt tatatagacg
     1861 gtcctcacgg gatggggaaa accaccacca cgcaactgct ggtggccctg ggttcgcgcg
     1921 acgatatcgt ctacgtaccc gagccgatga cttactggca ggtgctgggg gcttccgaga
     1981 caatcgcgaa catctacacc acacaacacc gcctcgacca gggtgagata tcggccgggg
     2041 acgcggcggt ggtaatgaca agcgcccaga taacaatggg catgccttat gccgtgaccg
     2101 acgccgttct ggctcctcat atcggggggg aggctgggag ctcacatgcc ccgcccccgg
     2161 ccctcaccct catcttcgac cgccatccca tcgccgccct cctgtgctac ccggccgcgc
     2221 gataccttat gggcagcatg accccccagg ccgtgctggc gttcgtggcc ctcatcccgc
     2281 cgaccttgcc cggcacaaac atcgtgttgg gggcccttcc ggaggacaga cacatcgacc
     2341 gcctggccaa acgccagcgc cccggcgagc ggcttgacct ggctatgctg gccgcgattc
     2401 gccgcgttta cgggctgctt gccaatacgg tgcggtatct gcagggcggc gggtcgtggc
     2461 gggaggattg gggacagctt tcggggacgg ccgtgccgcc ccagggtgcc gagccccaga
     2521 gcaacgcggg cccacgaccc catatcgggg acacgttatt taccctgttt cgggcccccg
     2581 agttgctggc ccccaacggc gacctgtaca acgtgtttgc ctgggccttg gacgtcttgg
     2641 ccaaacgcct ccgtcccatg cacgtcttta tcctggatta cgaccaatcg cccgccggct
     2701 gccgggacgc cctgctgcaa cttacctccg ggatggtcca gacccacgtc accacccccg
     2761 gctccatacc gacgatctgc gacctggcgc gcacgtttgc ccgggagatg ggggaggcta
     2821 actgaggatc ccacgtcact attgtatact ctatattata ctctatgtta tactctgtaa
     2881 tcctactcaa taaacgtgtc acgcctgtga aaccgtacta agtctcccgt gtattcttat
     2941 caccatcagg tgacatcctc gcccaggctg tcaatcatgc cggtatcgat tccagtagca
     3001 ccggccccac gctgacaacc cactcttgca gcgttagcag cgcccctctt aacaagccga
     3061 cccccaccag cgtcgcggtt actaacactc ctctccccga cctgcagccc agctagctac
     3121 acaagggagc caagagatga ccgaggctct tacacagcgt ccatctatca aaactatatg
     3181 gggaattctt tctctgggta ctcacactct cccccactac tgcaagtcaa ccgatcctcc
     3241 agcaactccc cgagaacgtc ggaaactgat gatggtgtgg taggaaagtc atctgatgga
     3301 gaagacgagc aacaggtccc caagggcccc atccattctc cagttgaact caagcatgtg
     3361 catgcaactg tggttaaagt tccagaagtg aattcctctg ccttgccaca caagctccgg
     3421 atcaaagcca aagccatgca gatcaaagta gaagcctttg ataatgaatt tgaggccacg
     3481 caaaaacttt cctcacctat tgacatgaca tctaaaagac atttcgaact cgaaaagcat
     3541 agtgccccaa gtatggtaca ttcttctctt actcctttct cagtgcaagt gactaacatt
     3601 caagattggt ctctcaaatc ggagcactgg catcaaaaag aactgagtgg caaaactcag
     3661 aatagtttca aaactggagt tgttgaaatg aaagacagtg gctacaaagt ttctgaccca
     3721 gagaacttgt atttgaagca ggggatagca aacttatctg cagaggttgt ctcactcaag
     3781 agacttatag ccacacaacc aatctctgct tcagactctg gggggagcgg aggaggttcc
     3841 ggtggaggtg gttctggaga ttacaaggat gacgacgata agggcgatta caaggatgac
     3901 gacgataagg gagattacaa ggatgacgac gataaggttt caggaagcgg agctactaac
     3961 ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctgg atcgtttcgc
     4021 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4081 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4141 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4201 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4261 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4321 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4381 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4441 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4501 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4561 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4621 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggccgaccg ctatcaggac
     4681 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4741 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4801 gacgagttct tctgattcga acatcgctgg gcatttagaa agatgtcatt tgcaatagag
     4861 cagtccattt tgtattatgc tgaattttca ctggacctgt gatgtcattt cactgtgatg
     4921 tgcacatgtt gtctgtttgg tgtctttttg tgcacagatt atgatgaaga ttagattgtg
     4981 ttatcactct gcctgtgtat agtcagatag tccatgcgaa ggctgtatat attgaacatt
     5041 atttttgttg ttctattata aagtgtgtaa gttaccagtt tcaataaagg attggtgaca
     5101 aacacagaac tcctgctcca ttgcatttga tttgatggag aagaaaagtc aatttaagtt
     5161 ataaaaataa atattaagat attttacatt ttatatgaga gagttttctc ccctgaataa
     5221 gagaaaactg aagtgctaag ttcagcaaat aatttgtttt tccaggcttc taaagaaaga
     5281 aaaataacag tgaattttct tgcatttgag actaattcac taaattactt tatattatat
     5341 atttttctgt aagattatgt tttctctata attttcttaa gtataaaaat acaagttgtt
     5401 tggtaacagc ctcttcttag ttatctaccc tttaagtatg acacatggat gtggatagtt
     5461 gagtcactga tgcattttat taaagtctgt tttaaaagag cagataaaga ctgaggatga
     5521 ggcattttta tgtcttggaa agtcctctcc actgactgta gcctccaatt cactggccgt
     5581 cgttcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5641 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5701 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gcgatttatt caacaaagcc
     5761 gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga
     5821 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaatata ttcatatcag gattatcaat
     5881 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     5941 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     6001 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     6061 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     6121 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     6181 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     6241 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     6301 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     6361 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     6421 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     6481 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     6541 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     6601 cttcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     6661 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     6721 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
     6781 tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat
     6841 caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct
     6901 ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac gaggcagacc
     6961 tctcgacgga
//
