LOCUS       sequence-492046-        5953 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  Insert a C-terminal HaloTag at GNT1 by popin-popout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5953)
  TITLE     Glick lab plasmids 2025
REFERENCE   2  (bases 1 to 5953)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: YIplac211-Gnt1-HaloTag
FEATURES             Location/Qualifiers
     source          1..5953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             860..1750
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(2451..2468)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2451..2467)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2460..2482)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2676..2695)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2795..2817
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2935..3738)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3739..3954)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3961..3979)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4047..4151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4152..4220,4221..5012)
                     /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(4370..4389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5183..5771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5672..5691
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5925..5942
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctgga
      241 cattataagt tctacatcat ctttatcgcc gttatttgat ttccaaagta gtaaaaaggt
      301 tggaaaatta aaatttgctt ctaatttaat ggttattaat ccttcaaagg aggcgtttga
      361 tgaaattgtg aacgttatgc tgcccaagat tttaaacaag aaggaaaagt acgatatgga
      421 tttgatcaat gaagaaatgt acaatttaaa gaagattatt tataaacagt ttattttctt
      481 cagaaaagtc cgaaaacttt tcaaaccaga agtattggtt ttgccatttg caaggtacgg
      541 cttactgacg ggttcattga gaaacccacg gcactattct ataatctata atgacgtttt
      601 gggttataag acattggata atgatggcaa cgatattccg gtaggattaa acgacagtgt
      661 cgcatattct aagtacatac atttttcgga ttatccattg gccaagccct ggaattatcc
      721 atcaatgaag gagtttgaat gtatcgttaa agaagaagac gcggaggatt caaagctaga
      781 gcaccaagct tgtgaccttt ggaactcggt gtacgcttct tacatacagt ccagggaaat
      841 ctgcctcgtt ggaagttcga tggcagaaat cggtacaggt ttcccttttg acccacacta
      901 tgtcgaagta ttaggtgaaa gaatgcacta tgttgatgtc ggtcctagag atggtactcc
      961 agttttgttt ttacatggta acccaacatc ttcatacgtt tggagaaaca tcatcccaca
     1021 tgttgcacca actcatagat gtattgctcc agatttgatt ggtatgggta aatctgataa
     1081 gccagatttg ggttatttct ttgatgatca tgttagattc atggatgctt ttattgaagc
     1141 attgggtttg gaagaagttg ttttggttat tcatgattgg ggttcagcat taggttttca
     1201 ttgggctaag agaaacccag aaagagttaa gggtatcgct tttatggaat tcattagacc
     1261 aattccaact tgggatgaat ggccagaatt cgcaagagaa acattccaag cttttagaac
     1321 tacagatgtt ggtagaaagt tgatcatcga tcaaaacgtt tttattgaag gtactttgcc
     1381 aatgggtgtt gttagaccat taacagaagt tgaaatggat cattacagag aaccattttt
     1441 aaacccagtt gatagagaac cattgtggag atttccaaat gaattaccaa ttgctggtga
     1501 accagcaaac atcgttgctt tggttgaaga atacatggat tggttacatc aatctccagt
     1561 tccaaagttg ttattttggg gtacaccagg tgttttgatt ccaccagcag aagctgcaag
     1621 attggctaag tcattgccaa actgtaaggc tgttgatatt ggtccaggtt taaatttgtt
     1681 acaagaagat aatccagact tgataggtag tgaaatagcc agatggttgt ccacattaga
     1741 aatctcaggt tgattatgag gagaagtgaa aattttcaaa aagtgtaaaa attagtatag
     1801 tcggaaacgc atatatatat atcatagtat tgataatgtt tctgccaaga ttgtgtccgt
     1861 tatcgcattt tctaacgggt aaggtatgta ctaggtgaga gcgggatctt atttcaacat
     1921 gttgtgcttt aatgcttctt tattcaacag cctgtcaacg tcatcaccgt attttgcgtt
     1981 accttttcca ttttctttga aggcataatc aacagtaatt ctgttgttgg cgactaactg
     2041 attgttcaaa gattttattg ccaaatctgc tttttcaaag tcctcaaaat aaacataagc
     2101 acactttaat tttccgtttg ataggtagaa aatttcgggt tcccttatga gcttaccaaa
     2161 tttattgaaa attttcacta attggtcgct atctatcgaa tctgcaaggt tttttatgaa
     2221 caacttagca ataggtagaa tcatgtcctt agaaatattt gaaggtaaat tagttgttcc
     2281 tgtagaattg gttacttgac gaacctttat cagtcggtcg tataatctaa cggtattatt
     2341 cattattttt attgcatatt gagcatctcc ttgattataa aattcgatga atgcgtagcc
     2401 ttggtaagct tgtagaacct agaggatccc cgggtaccga gctcgaattc actggccgtc
     2461 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2521 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2581 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     2641 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2701 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2761 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2821 tcaccgtcat caccgaaacg cgcgagacga aagggcgggt aataactgat ataattaaat
     2881 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     2941 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     3001 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     3061 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     3121 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     3181 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     3241 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     3301 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     3361 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     3421 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     3481 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     3541 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     3601 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     3661 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     3721 atatgtagct ttcgacatga tttatcttcg tttcctgcat gtttttgttc tgtgcagttg
     3781 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     3841 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     3901 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagctg
     3961 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4021 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4081 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4141 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4201 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4261 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4321 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4381 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4441 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4501 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4561 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4621 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4681 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4741 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4801 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4861 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4921 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4981 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5041 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5101 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5161 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5221 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5281 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5341 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5401 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5461 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5521 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5581 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5641 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5701 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5761 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5821 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5881 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5941 aggaagcgga aga
//
