LOCUS       sequence-194183-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human AHR.
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_AHR
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    412..433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(448..453,454..471,472..478)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    486..502
                     /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     491..513
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     510..526
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     510..526
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        703..905
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     856..876
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     902..926
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1002..2132
                     /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             3166..3189
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3193..3216
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3220..3243
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3259..3315
                     /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             3328..4122
                     /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(3382..3401)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3992..4011
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4893..4909)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4902..4924)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5128..5943)
                     /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     5851..5870
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5935..5954
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6323..6911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6812..6831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agaagctcgc acattcagca gcgtttttca gcgcgttttc gatcaacgtt tcaatgttgg
      181 tatcaacacc aggtttaact ttgaacttat cggcactgac ggttactgat tttgaacttt
      241 tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca
      301 gcaaaagttc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      361 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      421 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      481 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      541 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg aattagcttg
      601 catgcctgca ggtcaattcc ctggcattat gcccagtaca tgaccttatg ggactttcct
      661 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      721 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttcaagtctc caccccattg
      781 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      841 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      901 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
      961 ataggagaca ccgggaccga tccagcctgg ggatcccgac catggcttcg tacccctgcc
     1021 atcaacacgc gtctgcgttc gaccaggctg cgcgttctcg cggccatagc aaccgacgta
     1081 cggcgttgcg ccctcgccgg cagcaagaag ccacggaagt ccgcctggag cagaaaatgc
     1141 ccacgctact gcgggtttat atagacggtc ctcacgggat ggggaaaacc accaccacgc
     1201 aactgctggt ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag ccgatgactt
     1261 actggcaggt gctgggggct tccgagacaa tcgcgaacat ctacaccaca caacaccgcc
     1321 tcgaccaggg tgagatatcg gccggggacg cggcggtggt aatgacaagc gcccagataa
     1381 caatgggcat gccttatgcc gtgaccgacg ccgttctggc tcctcatatc gggggggagg
     1441 ctgggagctc acatgccccg cccccggccc tcaccctcat cttcgaccgc catcccatcg
     1501 ccgccctcct gtgctacccg gccgcgcgat accttatggg cagcatgacc ccccaggccg
     1561 tgctggcgtt cgtggccctc atcccgccga ccttgcccgg cacaaacatc gtgttggggg
     1621 cccttccgga ggacagacac atcgaccgcc tggccaaacg ccagcgcccc ggcgagcggc
     1681 ttgacctggc tatgctggcc gcgattcgcc gcgtttacgg gctgcttgcc aatacggtgc
     1741 ggtatctgca gggcggcggg tcgtggcggg aggattgggg acagctttcg gggacggccg
     1801 tgccgcccca gggtgccgag ccccagagca acgcgggccc acgaccccat atcggggaca
     1861 cgttatttac cctgtttcgg gcccccgagt tgctggcccc caacggcgac ctgtacaacg
     1921 tgtttgcctg ggccttggac gtcttggcca aacgcctccg tcccatgcac gtctttatcc
     1981 tggattacga ccaatcgccc gccggctgcc gggacgccct gctgcaactt acctccggga
     2041 tggtccagac ccacgtcacc acccccggct ccataccgac gatctgcgac ctggcgcgca
     2101 cgtttgcccg ggagatgggg gaggctaact gaggatccca cgtcactatt gtatactcta
     2161 tattatactc tatgttatac tctgtaatcc tactcaataa acgtgtcacg cctgtgaaac
     2221 cgtactaagt ctcccgtgta ttcttatcac catcaggtga catcctcgcc caggctgtca
     2281 atcatgccgg tatcgattcc agtagcaccg gccccacgct gacaacccac tcttgcagcg
     2341 ttagcagcgc ccctcttaac aagccgaccc ccaccagcgt cgcggttact aacactcctc
     2401 tccccgacct gcagcccagc tctatgcttt cagtagtatt cattaattaa aagtatatta
     2461 tgaataagaa atcggttaag aaaatcatac ctgtggtgta atattttagg aatatttaga
     2521 aacatttaga aaatcagaag ctgccttttt tggaaaagtt ttgcattcca gtgtaagatt
     2581 cctgaatata atcaattcat tgatacctat aacttcaaac ctggtcaaag caaacctact
     2641 tatgatagag aatatttaac tgtaatttgt gacttgtgct gtttgaccca tcaattccta
     2701 cctgacccaa aggcaatctg aaatgagaca gctgcatcac tcatctgtac catgttcaaa
     2761 atttcctgtc aaattaatgg aaaaacaggt tataaatgca actaatacat aatatgtcct
     2821 cagtatgtct tcactgtgtg aaagatttaa aatttagcaa cagtaaagga acttgaagtt
     2881 tacaactctc aggggtaaga ttttaaaaat acatgttaat gttatttact ggcttaagat
     2941 acttggaaga tctattccaa taagttgcat caccattttt gttttcagtt tcagaatgga
     3001 gttttaaatg aaacatatcc agctgaatta aataacataa ataacactca gactaccaca
     3061 catcttcagc cacttcatca tccgtcagaa gccagacctt ttcctgattt gacatccagt
     3121 ggattcctgg ggagcggagg aggttccggt ggaggtggtt ctggagatta caaggatgac
     3181 gacgataagg gcgattacaa ggatgacgac gataagggag attacaagga tgacgacgat
     3241 aaggtttcag gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag
     3301 gagaaccctg gacctggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3361 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3421 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3481 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3541 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3601 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3661 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3721 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3781 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3841 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3901 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3961 ctgggtgtgg ccgaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4021 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4081 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gattcgaaca tctttgaccc
     4141 tggtttttgg attaaattag tttgtgaagg attatggaaa aataaaactg tcactgttgg
     4201 acgtcagcaa gttcacatgg aggcattgat gcatgctatt cacaattatt ccaaaccaaa
     4261 ttttaatttt tgcttttaga aaagggagtt taaaaatggt atcaaaatta catatactac
     4321 agtcaagata gaaagggtgc tgccacggag tggtgaggta ccgtctacat ttcacattat
     4381 tctgggcacc acaaaatata caaaacttta tcagggaaac taagattctt ttaaattaga
     4441 aaatattctc tatttgaatt atttctgtca cagtaaaaat aaaatacttt gagttttgag
     4501 ctactggatt cttattagtt ccccaaatac aaagttagag aactaaacta gtttttccta
     4561 tcatgttaac ctctgctttt atctcagatg ttaaaataaa tggtttggtg ctttttataa
     4621 aaagataatc tcagtgcttt cctccttcac tgtttcatct aagtgcctca catttttttc
     4681 tacctataac actctaggat gtatatttta tataaagtat tctttttctt ttttaaatta
     4741 atatctttct gcacacaaat attatttgtg tttcctaaat ccaaccattt tcattaattc
     4801 aggcatattt taactccact gcttacctac tttcttcagg cttggaaagt cctctccact
     4861 gactgtagcc tccaattcac tggccgtcgt tcactggccg tcgttttaca acgtcgtgac
     4921 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4981 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5041 ggcgaatgcg atttattcaa caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca
     5101 gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg
     5161 caatatattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     5221 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     5281 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     5341 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat
     5401 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     5461 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     5521 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     5581 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     5641 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     5701 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     5761 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     5821 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     5881 agcatccatg ttggaattta atcgcggctt cgagcaagac gtttcccgtt gaatatggct
     5941 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     6001 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat
     6061 aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg
     6121 ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat
     6181 caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc
     6241 agcaacacct tcttcacgag gcagacctct cgacggagtt ccactgagcg tcagaccccg
     6301 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6361 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6421 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6481 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6541 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6601 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6661 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6721 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6781 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6841 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6901 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6961 ttgctcacat
//
