LOCUS       sequence-454189-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blast-T2A-2xHA-FKPB12(dTagDegron) mouse Nipbl N-term targeting 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  AUTHORS   Hansen KL, Adachi AS, Braccioli L, Kadvani S, Boileau RM, Martinovic
            M, Pokorny B, Shah R, Anderson EC, Zhang K, Carel I, Bonitto K, 
            Blelloch R, Fudenberg G, de Wit E, Nora EP
  TITLE     Synergy between regulatory elements can render cohesin dispensable 
            for distal enhancer function.
  JOURNAL   Science. 2025 Nov 27:eadt4221. doi: 10.1126/science.adt4221.
  PUBMED    41308125
REFERENCE   2  (bases 1 to 5378)
  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: "Science. 
            2025 Nov 27:eadt4221. doi: 10.1126/science.adt4221."
COMMENT     Sequence Label: pEA038
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1364..1759
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1775..1828
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1835..1861
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1865..1891
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1865..1891
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1898..2218
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(3157..3173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3157..3173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3170..3192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3181..3197
                     /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(3205..3211,3212..3229,3230..3235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3250..3271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3388..3405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3559..4147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3639..3658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4318..5109,5110..5178))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4941..4960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5179..5283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5351..5369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagttcgcg agaaacaggg gaagatggac
      421 aggcgagata gctcagcaag gaaaagcacc aaccacaagc ctgagctgct gagtacagtc
      481 ctccagactc ttgtgctgag ggaaagatcc tagttccgca gattatcttc tgatctgcac
      541 acacatcagt catagcaggc acacaacaat acacgtggac actaagtgaa caaatagtaa
      601 caaattacaa atggggcagg ggaggaggag ggacatatca agtgtgtcac aaaggaatat
      661 acagcaattt actgagaaaa tattgggcaa taaatgtcat aaattcagtt atcctaaggt
      721 aaaatggctg tggaatgaaa gtgttcttga attgtaggat ttagaagaga ggagaataaa
      781 agtctcacac tgcaaaccac aaggatccaa aagaatctga agagatactc ttggtaggag
      841 gggcaagtcc atttagagag taaaaaggga actagacaag aaattgaagc cagattatta
      901 agtaaaaatt ttattgtatt tagagtttag aaagagtacc ctgccctagc agtatataca
      961 ccagaagtta agaaacaaga ttctggagct agattgtgtc attagaatta cagctccagt
     1021 agttacttta aactcactgt aattttctca ttagtaaaat gcagaaaaga gtaatactct
     1081 gctgtacctg ggttgttgtg agacctaaat cttaatgtat acaaagcatt ttgcagtgtt
     1141 taatgtatgg cacatgcttg ttaaacacat tgaacaaata gaagctaaga gttttgtttc
     1201 taaatgaata agtatctctc tgctactcat atatttgaca aagttctaca aataattata
     1261 tggttttctt tgtgtgtgtt gcagagtttg ggaaatggaa agtaatgaca gctggcacct
     1321 gaactaaata ctttcataga caacaccatt ccagaacttc aggatggcca agcctttgtc
     1381 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     1441 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     1501 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     1561 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     1621 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     1681 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     1741 tggttatgtg tgggagggcg ctagcggcag tggagagggc agaggaagtc tgctaacatg
     1801 cggtgacgtc gaggagaatc ctggcccagg tacctaccca tacgacgtac ccgattatgc
     1861 tggctacccc tacgacgtgc ccgactacgc cgccggcgga gtgcaggtgg aaaccatctc
     1921 cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg
     1981 gatgcttgaa gatggaaaga aagttgattc ctcccgggac agaaacaagc cctttaagtt
     2041 tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt
     2101 gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc
     2161 aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaaag
     2221 tggtggcggt ggctcgggcg gtggtgggtc gggtggctcc ggaatgaatg gggatatgcc
     2281 ccatgtcccc ataactacgc ttgcggggat tgctagtctt acggaccgta agtttgcata
     2341 attttcctaa gttctttact ttgctaatac tcatttttag agacaaaaaa ttacctttaa
     2401 tgtcacatat aacagaaaat atgtatttga ttagtgtggg gggagtagca aaattaatgt
     2461 ttccacctaa tacataaatt taatttcttc atccttctgt gaactcctaa agaaatgata
     2521 gtgtgataat ttttataaag accttaaaaa tcaataagga ctttagaaaa atgacacaaa
     2581 gctttaagag atgttaaaac ttaaataaac atgggttaat aaacaaaact atggtctttt
     2641 ttttaagaaa gcgtttacaa aaacagtacc tgaaagtgtc ctccaagaat gagaaattga
     2701 tcgcccacat tacctaaact gtagtggtcc tgattttttt aatatcaggt tgaattgtta
     2761 gttagaattt gttaacaggt atgagatatg tcatcactat gagagcccac tttactctga
     2821 acactaaggt attaaaaatt attgtcaata cttagaaatt tagttaatag aagttttaag
     2881 gttcatattt tggtagacac ataactattc ttttccttgg gtaatggcac agagtctgcc
     2941 caagtgtttg tccctatgtc atcctccttt tagtatagga atagagataa aggaaaggcc
     3001 agaaaagaaa aaacaaaatg aaaacataaa tacctttttc ttatagcagt atgtaagata
     3061 tggaactttg caataatcta tgaaaaaata acttccagaa ctgttttcaa ctgtgctcag
     3121 aatccctccc tgggttttat gcatggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5281 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5341 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
