LOCUS       sequence-423373-        8504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human BCL11A. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8504)
  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 8504)
  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: pHTC-HaloTag_BCL11A:1-773
FEATURES             Location/Qualifiers
     source          1..8504
                     /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"
     CDS             1191..1202
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3458..3478
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3494..4378
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(4450..4468)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4505..4626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4516..4535)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4570..4589
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4871..5228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4871..5062
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4894..4914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5079..5214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5279..6073
                     /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(5333..5352)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5943..5962
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6137..6185
                     /note="synthetic polyadenylation signal"
     promoter        6341..6445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6446..6514,6515..7306)
                     /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(6664..6683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7464..8052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7544..7563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8168..8451
                     /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 gcgatcgctt ccgaattcgc
     1081 caccatgtct cgccgcaagc aaggcaaacc ccagcactta agcaaacggg aattctcgcc
     1141 cgagcctctt gaagccattc ttacagatga tgaaccagac cacggcccgt tgggagctcc
     1201 agaaggggat catgacctcc tcacctgtgg gcagtgccag atgaacttcc cattggggga
     1261 cattcttatt tttatcgagc acaaacggaa acaatgcaat ggcagcctct gcttagaaaa
     1321 agctgtggat aagccacctt ccccttcacc aatcgagatg aaaaaagcat ccaatcccgt
     1381 ggaggttggc atccaggtca cgccagagga tgacgattgt ttatcaacgt catctagagg
     1441 aatttgcccc aaacaggaac acatagcaga taaacttctg cactggaggg gcctctcctc
     1501 ccctcgttct gcacatggag ctctaatccc cacgcctggg atgagtgcag aatatgcccc
     1561 gcagggtatt tgtaaagatg agcccagcag ctacacatgt acaacttgca aacagccatt
     1621 caccagtgca tggtttctct tgcaacacgc acagaacact catggattaa gaatctactt
     1681 agaaagcgaa cacggaagtc ccctgacccc gcgggttggt atcccttcag gactaggtgc
     1741 agaatgtcct tcccagccac ctctccatgg gattcatatt gcagacaata acccctttaa
     1801 cctgctaaga ataccaggat cagtatcgag agaggcttcc ggcctggcag aagggcgctt
     1861 tccacccact ccccccctgt ttagtccacc accgagacat cacttggacc cccaccgcat
     1921 agagcgcctg ggggcggaag agatggccct ggccacccat cacccgagtg cctttgacag
     1981 ggtgctgcgg ttgaatccaa tggctatgga gcctcccgcc atggatttct ctaggagact
     2041 tagagagctg gcagggaaca cgtctagccc accgctgtcc ccaggccggc ccagccctat
     2101 gcaaaggtta ctgcaaccat tccagccagg tagcaagccg cccttcctgg cgacgccccc
     2161 cctccctcct ctgcaatccg cccctcctcc ctcccagccc ccggtcaagt ccaagtcatg
     2221 cgagttctgc ggcaagacgt tcaaatttca gagcaacctg gtggtgcacc ggcgcagcca
     2281 cacgggcgag aagccctaca agtgcaacct gtgcgaccac gcgtgcaccc aggccagcaa
     2341 gctgaagcgc cacatgaaga cgcacatgca caaatcgtcc cccatgacgg tcaagtccga
     2401 cgacggtctc tccaccgcca gctccccgga acccggcacc agcgacttgg tgggcagcgc
     2461 cagcagcgcg ctcaagtccg tggtggccaa gttcaagagc gagaacgacc ccaacctgat
     2521 cccggagaac ggggacgagg aggaagagga ggacgacgag gaagaggaag aagaggagga
     2581 agaggaggag gaggagctga cggagagcga gagggtggac tacggcttcg ggctgagcct
     2641 ggaggcggcg cgccaccacg agaacagctc gcggggcgcg gtcgtgggcg tgggcgacga
     2701 gagccgcgcc ctgcccgacg tcatgcaggg catggtgctc agctccatgc agcacttcag
     2761 cgaggccttc caccaggtcc tgggcgagaa gcataagcgc ggccacctgg ccgaggccga
     2821 gggccacagg gacacttgcg acgaagactc ggtggccggc gagtcggacc gcatagacga
     2881 tggcactgtt aatggccgcg gctgctcccc gggcgagtcg gcctcggggg gcctgtccaa
     2941 aaagctgctg ctgggcagcc ccagctcgct gagccccttc tctaagcgca tcaagctcga
     3001 gaaggagttc gacctgcccc cggccgcgat gcccaacacg gagaacgtgt actcgcagtg
     3061 gctcgccggc tacgcggcct ccaggcagct caaagatccc ttccttagct tcggagactc
     3121 cagacaatcg ccttttgcct cctcgtcgga gcactcctcg gagaacggga gcttgcgctt
     3181 ctccacaccg cccggggagc tggacggagg gatctcgggg cgcagcggca cgggaagtgg
     3241 agggagcacg ccccatatta gtggtccggg cccgggcagg cccagctcaa aagagggcag
     3301 acgcagcgac acttgttctt cacacacccc cattcggcgt agtacccaga gagctcaaga
     3361 tgtgtggcag ttttcggatg gaagctcgag agcccttaag ttccggatat ctagatttgg
     3421 gcccaattcc tgcaggcgag ctctcgagcc aaccactgag gatctgtact ttcagagcga
     3481 taacgatgga tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct
     3541 gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct
     3601 gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac
     3661 ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg
     3721 ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga
     3781 agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg
     3841 caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg
     3901 ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg
     3961 ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt
     4021 ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga
     4081 ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat
     4141 cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct
     4201 gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag
     4261 cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa
     4321 cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccggtta
     4381 atagaattgg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     4441 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     4501 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     4561 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4621 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     4681 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt
     4741 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4801 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     4861 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4981 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5041 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5101 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5161 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5221 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat
     5281 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5341 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5401 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5461 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5521 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5581 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5641 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5701 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5761 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5821 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5881 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5941 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6001 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6061 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6121 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     6181 gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc
     6241 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtaat
     6301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca ctttcaaagt
     6661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6841 ggccaactta cttctgacaa ctatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6961 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7021 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7081 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7141 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7201 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7261 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaattcg aaatgaccga
     7321 ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7381 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7441 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7501 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7561 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7621 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7681 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7741 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7801 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7861 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7921 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7981 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8041 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8101 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8161 ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca
     8221 tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc
     8281 ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt
     8341 gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac
     8401 aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg
     8461 cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
