LOCUS       sequence-423352-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human BCL11A. N-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 8451)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pHTN-HaloTag_BCL11A:1-773
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2121..2132
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4402..4420)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4452..4573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4463..4482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4517..4536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4818..5175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4818..5009
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4841..4861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5026..5161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5226..6020
                     /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(5280..5299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5890..5909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6084..6132
                     /note="synthetic polyadenylation signal"
     promoter        6288..6392
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6393..6461,6462..7253)
                     /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     complement(6611..6630)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7411..7999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7491..7510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8115..8398
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgcttccga attcatgtct cgccgcaagc aaggcaaacc
     2041 ccagcactta agcaaacggg aattctcgcc cgagcctctt gaagccattc ttacagatga
     2101 tgaaccagac cacggcccgt tgggagctcc agaaggggat catgacctcc tcacctgtgg
     2161 gcagtgccag atgaacttcc cattggggga cattcttatt tttatcgagc acaaacggaa
     2221 acaatgcaat ggcagcctct gcttagaaaa agctgtggat aagccacctt ccccttcacc
     2281 aatcgagatg aaaaaagcat ccaatcccgt ggaggttggc atccaggtca cgccagagga
     2341 tgacgattgt ttatcaacgt catctagagg aatttgcccc aaacaggaac acatagcaga
     2401 taaacttctg cactggaggg gcctctcctc ccctcgttct gcacatggag ctctaatccc
     2461 cacgcctggg atgagtgcag aatatgcccc gcagggtatt tgtaaagatg agcccagcag
     2521 ctacacatgt acaacttgca aacagccatt caccagtgca tggtttctct tgcaacacgc
     2581 acagaacact catggattaa gaatctactt agaaagcgaa cacggaagtc ccctgacccc
     2641 gcgggttggt atcccttcag gactaggtgc agaatgtcct tcccagccac ctctccatgg
     2701 gattcatatt gcagacaata acccctttaa cctgctaaga ataccaggat cagtatcgag
     2761 agaggcttcc ggcctggcag aagggcgctt tccacccact ccccccctgt ttagtccacc
     2821 accgagacat cacttggacc cccaccgcat agagcgcctg ggggcggaag agatggccct
     2881 ggccacccat cacccgagtg cctttgacag ggtgctgcgg ttgaatccaa tggctatgga
     2941 gcctcccgcc atggatttct ctaggagact tagagagctg gcagggaaca cgtctagccc
     3001 accgctgtcc ccaggccggc ccagccctat gcaaaggtta ctgcaaccat tccagccagg
     3061 tagcaagccg cccttcctgg cgacgccccc cctccctcct ctgcaatccg cccctcctcc
     3121 ctcccagccc ccggtcaagt ccaagtcatg cgagttctgc ggcaagacgt tcaaatttca
     3181 gagcaacctg gtggtgcacc ggcgcagcca cacgggcgag aagccctaca agtgcaacct
     3241 gtgcgaccac gcgtgcaccc aggccagcaa gctgaagcgc cacatgaaga cgcacatgca
     3301 caaatcgtcc cccatgacgg tcaagtccga cgacggtctc tccaccgcca gctccccgga
     3361 acccggcacc agcgacttgg tgggcagcgc cagcagcgcg ctcaagtccg tggtggccaa
     3421 gttcaagagc gagaacgacc ccaacctgat cccggagaac ggggacgagg aggaagagga
     3481 ggacgacgag gaagaggaag aagaggagga agaggaggag gaggagctga cggagagcga
     3541 gagggtggac tacggcttcg ggctgagcct ggaggcggcg cgccaccacg agaacagctc
     3601 gcggggcgcg gtcgtgggcg tgggcgacga gagccgcgcc ctgcccgacg tcatgcaggg
     3661 catggtgctc agctccatgc agcacttcag cgaggccttc caccaggtcc tgggcgagaa
     3721 gcataagcgc ggccacctgg ccgaggccga gggccacagg gacacttgcg acgaagactc
     3781 ggtggccggc gagtcggacc gcatagacga tggcactgtt aatggccgcg gctgctcccc
     3841 gggcgagtcg gcctcggggg gcctgtccaa aaagctgctg ctgggcagcc ccagctcgct
     3901 gagccccttc tctaagcgca tcaagctcga gaaggagttc gacctgcccc cggccgcgat
     3961 gcccaacacg gagaacgtgt actcgcagtg gctcgccggc tacgcggcct ccaggcagct
     4021 caaagatccc ttccttagct tcggagactc cagacaatcg ccttttgcct cctcgtcgga
     4081 gcactcctcg gagaacggga gcttgcgctt ctccacaccg cccggggagc tggacggagg
     4141 gatctcgggg cgcagcggca cgggaagtgg agggagcacg ccccatatta gtggtccggg
     4201 cccgggcagg cccagctcaa aagagggcag acgcagcgac acttgttctt cacacacccc
     4261 cattcggcgt agtacccaga gagctcaaga tgtgtggcag ttttcggatg gaagctcgag
     4321 agcccttaag ttctgagata tctagatttg ggcccaattc ctgcaggatt ttgcggccgc
     4381 ttgctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggccgcttcg
     4441 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4501 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4561 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     4621 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     4681 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4741 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     4801 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5041 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5101 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5161 ctaggctttt gcaaaaagct tgattcttct gacacaacag tctcgaactt aaggctagag
     5221 ccaccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5281 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5341 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5401 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5461 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5521 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5581 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5641 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5701 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5761 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5821 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5881 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5941 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6001 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6061 caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     6121 gttttttgtg tgaatcgata gcgataagga tcctctttgc gcttgcgttt tcccttgtcc
     6181 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     6241 acgtaatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6301 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6361 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6421 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6481 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6541 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6601 tcaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6661 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6721 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6781 acactgcggc caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt
     6841 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6901 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6961 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7021 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7081 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7141 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7201 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa
     7261 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     7321 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7381 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7441 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7501 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7561 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7621 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7681 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7741 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7801 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7861 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7921 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7981 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8041 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8101 gatcctttta tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt
     8161 gaaaccatga aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc
     8221 tggttgcata ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg
     8281 gtggtttgtt gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg
     8341 tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga
     8401 taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
