LOCUS       sequence-264347-        5565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and HaloTag-linker sequence for N-terminus tagging of 
            human UBTF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5565)
  TITLE     Allen Institute for Cell Sciences Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5565)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-80:UBTF-HaloTag
FEATURES             Location/Qualifiers
     source          1..5565
                     /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             1413..2303
                     /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             2906..2917
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3344..3360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3344..3360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3357..3379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3368..3384
                     /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(3392..3398,3399..3416,3417..3422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3437..3458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3575..3592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3746..4334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3826..3845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4505..5296,5297..5365))
                     /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     5128..5147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5366..5470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5538..5556
                     /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 ccagtgaatt gacgcgtatt gggatgcttc
      421 tcggagccgg caggcgcagt tctccccggc cgccggaggg ggcggcgggg ccggtgagcg
      481 ctgcgccccc gcctcccggc cgccgtcctc ccgccctctc cttggctcgg ctccccggag
      541 gccggggcgg gggcgccggc ggggctgggc cgcaggctcg ggggaggagg cggcggttcc
      601 cgccgcggtc gcgcctctgt gccgggctcc tccgtgcgcg tcggggtggc tgggcggtgg
      661 cggcgcctct ggccggggtg ggtggctcgg gggcgggggg caggctaggc cgcttctctc
      721 cggctgcttg cccgcctgtg cgggcgggga cggcgaaggg agcccgggag gggggagcgg
      781 cgggaatccg gatccggtcc ccgggggcgg tgcggccgct ccgctccggg gctggcggcg
      841 gctgcgctgc gtccgggtcc ccgccgggtt gcggaggcgg ggggaaccgg ccggggggag
      901 gcgggaggag gagactgggg cgcgtgcccg cttccgtggc cgcgcgcgcc ctttacccgc
      961 ccttcctctt cccccgccct ccagcctggg gcctccccgg ggcgggcgct ccgaggagga
     1021 ggagattttt tctttttttt tttttttggc tggggttggg aagcggggcg gagggcgggc
     1081 gctcgctgca cgcgggagct aggagcaggc cctcgcgtgg ccaggggagg gcgcgcgccg
     1141 cggagccggg ctgcaggggc tgggggaggg ggtcacgagg aagaggaggg gcgcactccc
     1201 attcccatag accccttaag tctaataagg tcatggctga gtctctcaga gtgtggacct
     1261 gcccccttct actctgggcg gttagaaagt cgttagtcgt tcacctctca gccttcctga
     1321 gtgggaactg gtatgtcttc caaagggtga ggtggctttg accccgggtt gcccggccag
     1381 cacgaccgag gaggtggctg gacagctaga ggatggcaga aatcggtact ggctttccat
     1441 tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc
     1501 gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca
     1561 acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg
     1621 gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg
     1681 ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg
     1741 ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg
     1801 agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc
     1861 aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg
     1921 agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc
     1981 gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc
     2041 caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc
     2101 accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg
     2161 ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg
     2221 gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc
     2281 tgtcgacgct cgagatttcc ggctccggca acggagaagc cgactgcccc accgatctgg
     2341 aaatggccgc ccccaaaggc caaggtaggg gcatccaccc ccggccctct gaaacttaaa
     2401 ccctctttcc ctcgtggctg cggggctcac tcccttcttc aagggctttt tcagtctgca
     2461 tacactgaac tgggggagaa agtgccttga gtatttatga atcaagcatc tcccacactt
     2521 ggggtttggc tgaatccaaa ttatttggcc tggagtggct cacctcccct ctctccctgt
     2581 ccctcaccct ctgtgttaga ggaggcctgc cttctggggt tcccattcag cctgaagtgt
     2641 gtcccagccc agtgtcctgg aatgggagga aggatgagtg tagaggaagg gtcaggaagg
     2701 ggttggggaa ggcttctcag gctttttagg aagaccatga ggcttttgta gaggcaggtg
     2761 gagaaaaggg ctgttttggt ggttttaggc cccaagccat gggatatatg tactcaacta
     2821 gatctctgcc tcattcagca gtgatccagt ctagatgggg atagagagta ggatgaagct
     2881 ggacagctgg ccatgcttgt gggacattga ggggagaggg gattatgggt ccacgtggcc
     2941 cattctgcag aatctctggc ccaggttacc agatgcaaat gaaaatggag agcccttttg
     3001 acttctggcc ttgggtggct attctcccag cccatccaca gtggcctaga gctctctggg
     3061 gctgcatttc agtggctgaa aaataactgt tccagacctg ttcttgcttc cctgggtagt
     3121 tgggctgaca agacagcttg taattctggg ggttctccct ggagcagggc cctgagagtt
     3181 tctgtgcacc tgcctacctt gcctcagagc tgccctgtat cctgcttctg cttctgtttc
     3241 ctttttctta tttgccagaa attctccctt gatctgcttt aaggggctag ggggtggtgg
     3301 gtgtctctaa tcccaatggc gcgccgagct tggcgtaatc atggtcatag ctgtttcctg
     3361 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3421 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3481 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3541 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3601 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3661 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3721 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3781 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3841 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3901 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3961 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4021 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4081 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4141 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4201 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4261 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4321 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4381 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4441 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4501 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4561 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4621 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4681 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4741 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4801 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4861 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4921 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4981 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5041 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5101 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5161 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5221 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5281 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5341 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5401 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5461 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5521 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
