LOCUS       sequence-173001-        5059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tag S. pombe genes in C terminal with HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5059)
  TITLE     pFA6a-mEGFP-KanMX6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5059)
  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            "Unpublished"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: pFA6a-HaloTag-KanMX6
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..950
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     terminator      974..1161
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1029..1053)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1236..2592
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1236..1579
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1580..2389
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTSIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1647..1666)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2395..2592
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2696..2715)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2697..2715)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2802..2819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2973..3561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3053..3072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3732..4523,4524..4592))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITISDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4355..4374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4593..4697)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4765..4783
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4821..4843)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4943..4962
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5043..5059,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5043..5059,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acgcagaaat cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc
      121 gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta
      181 acccgacctc ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct
      241 gcattgctcc agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct
      301 tcgacgacca cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg
      361 tcctggtcat tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag
      421 agcgcgtcaa aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat
      481 ggccagaatt tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc
      541 tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc
      601 tgactgaagt cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc
      661 cactgtggcg cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc
      721 tggtcgaaga atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg
      781 gcaccccagg cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta
      841 actgcaaggc tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc
      901 tgatcggcag cgagatcgcg cgctggctgt cgacgctcga gatttccggc tagggcgcgc
      961 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     1021 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     1081 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct
     1141 tatttaccac acctctaccg gcagatccgc tagggataac agggtaatat agatctgttt
     1201 agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc
     1261 ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc
     1321 ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa
     1381 gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg
     1441 cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac
     1501 tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca
     1561 catccgaaca taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa
     1621 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1681 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1741 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1801 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1861 actcaccact tcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc
     1921 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1981 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2041 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2101 acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca
     2161 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2221 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2281 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2341 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac
     2401 aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt
     2461 gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc
     2521 ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg
     2581 gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat
     2641 tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta
     2701 tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc
     2761 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2821 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggctgt aatacggtta
     2881 tccacagaat caggggataa cgcaggaaag aacatgttag caaaaggcca gcaaaaggcc
     2941 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3001 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3061 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3121 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3181 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3241 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3301 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3361 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3421 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3481 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3541 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3601 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3661 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3721 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3781 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3841 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3901 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3961 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4021 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4081 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4141 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4201 gtgttatcac taatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4261 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4321 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4381 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4441 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4501 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4561 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4621 atttatcagg gttattgtct cattagcgga tacatatttg aatgtattta gaaaaataaa
     4681 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4741 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4801 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4861 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4921 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4981 gacatattgt cgttataacg cggctacaat taatacataa ccttatgtat catacacata
     5041 cgatttaggt gacactata
//
