LOCUS       sequence-492076-        7117 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  Insert a C-terminal HaloTagx2 at SYS1 by popin-popout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7117)
  TITLE     Glick lab plasmids 2025
REFERENCE   2  (bases 1 to 7117)
  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-Sys1-HaloTagx2
FEATURES             Location/Qualifiers
     source          1..7117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(942..1733,1734..1802))
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2000..2215
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2216..3019
                     /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"
     primer_bind     complement(3137..3159)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3259..3278
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3472..3494
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3486..3503
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3487..3503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4563..5453
                     /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             5460..6350
                     /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(6898..6914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6898..6914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6911..6933)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6922..6938
                     /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(6946..6952,6953..6970,6971..6976))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6991..7012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccagcttt tcaattcatc attttttttt tattcttttt tttgatttcg
     2041 gtttccttga aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga
     2101 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     2161 agtattctta acccaactgc acagaacaaa aacatgcagg aaacgaagat aaatcatgtc
     2221 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     2281 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     2341 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     2401 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     2461 caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa
     2521 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     2581 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa
     2641 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg
     2701 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
     2761 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     2821 cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa ccgtggatga
     2881 tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     2941 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     3001 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     3061 aattagagct tcaatttaat tatatcagtt attacccgcc ctttcgtctc gcgcgtttcg
     3121 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3181 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3241 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     3301 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     3361 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3421 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3481 gacgttgtaa aacgacggcc agtgaattcc tggtaatatt tcagttgttg actaatatat
     3541 atagtggttt ttgccaaaat taaagacttc cttttattaa ctaggtgacc agatataagg
     3601 aagtgtattt gctttctaat aacttgtgta cggccataac ccggctttta atcagtttat
     3661 ttaacaaggt atacaaatcg ccaaacgtaa caagagatag gcgctattat tcatttaata
     3721 ttgtcgtatt caaaatacta acggtgaaca atttgctcca ttgtaacact ataggacaag
     3781 gctaatcatg aataaatatc tccccaccca aaatattcgt agctcgcgat attcacggta
     3841 atattctgta acttcatgtt aagagatgat ctttaggaag gctaaggaag ggtacacata
     3901 gaagtgagat aaaaaatagg catttgaatt actgtaacat ggtttcgata agaaggtatt
     3961 tgcgagtgcc gaatgagttg aaaccttccc agatcttcaa acaagattct ctctctccaa
     4021 gtaagattgg acttcaaatt gttcttttac agatttttta ttacaccaca gcgatcgttc
     4081 ttttttattg ttgggcgaag ctagcgggat atgatctcaa tatcaaggaa tggctgtttt
     4141 cttgggagaa tatcgacttt acgaacgcat acggtttatc aatctcactg ttatggcttc
     4201 tggattcatt aatatgtgtt tttttcttaa ctgttatcgt tggacggagt aaactcgcat
     4261 gggattttgc tatcactata catgccatca attttattgt ggtgttcctc tatactcgga
     4321 aatttccttc tttttcttgg ttttttcttc aaattttatc atctttaata ctaatatttt
     4381 tggggacatg gacaacaaga tggagagagc ttagagacac cttttttgag ggcttggttg
     4441 atcctaatga aggagaagtt ggattggtta cgcctagtca gcagcatagt aatcattcag
     4501 aattggagca atcaccaata caactaaaag acttagaaag ccaaatacgg gatccaacta
     4561 gtatggctga aatcggtaca ggttttcctt ttgacccaca ctatgtcgaa gtattaggtg
     4621 aaagaatgca ctatgttgat gtcggtccta gagatggtac tccagttttg tttttacatg
     4681 gtaacccaac atcttcatac gtttggagaa acatcatccc acatgttgca ccaactcata
     4741 gatgtattgc tccagatttg attggtatgg gtaaatctga taagccagat ttgggttatt
     4801 tctttgatga tcatgttaga ttcatggatg cttttattga agcattgggt ttggaagaag
     4861 ttgttttggt tattcatgat tggggttcag cattaggttt tcattgggct aagagaaacc
     4921 cagaaagagt taagggtatc gcttttatgg aattcattag accaattcca acttgggatg
     4981 aatggccaga attcgcaaga gaaacattcc aagcttttag aactacagat gttggtagaa
     5041 agttgatcat cgatcaaaac gtttttattg aaggtacttt gccaatgggt gttgttagac
     5101 cattaacaga agttgaaatg gatcattaca gagaaccatt tttaaaccca gttgatagag
     5161 aaccattgtg gagatttcca aatgaattac caattgctgg tgaaccagca aacatcgttg
     5221 ctttggttga agaatacatg gattggttac atcaatctcc agttccaaag ttgttatttt
     5281 ggggtacacc aggtgttttg attccaccag cagaagctgc aagattggct aagtcattgc
     5341 caaactgtaa ggctgttgat attggtccag gtttaaattt gttacaagaa gataatccag
     5401 acttaatagg ttccgaaatc gccagatggt tatcaacttt agaaatcagt ggtgctagta
     5461 tggctgaaat cggtacaggt tttccttttg acccacacta tgtcgaagta ttaggtgaaa
     5521 gaatgcacta tgttgatgtc ggtcctagag atggtactcc agttttgttt ttacatggta
     5581 acccaacatc ttcatacgtt tggagaaaca tcatcccaca tgttgcacca actcatagat
     5641 gtattgctcc agatttgatt ggtatgggta aatctgataa gccagatttg ggttatttct
     5701 ttgatgatca tgttagattc atggatgctt ttattgaagc attgggtttg gaagaagttg
     5761 ttttggttat tcatgattgg ggttcagcat taggttttca ttgggctaag agaaacccag
     5821 aaagagttaa gggtatcgct tttatggaat tcattagacc aattccaact tgggatgaat
     5881 ggccagaatt cgcaagagaa acattccaag cttttagaac tacagatgtt ggtagaaagt
     5941 tgatcatcga tcaaaacgtt tttattgaag gtactttgcc aatgggtgtt gttagaccat
     6001 taacagaagt tgaaatggat cattacagag aaccattttt aaacccagtt gatagagaac
     6061 cattgtggag atttccaaat gaattaccaa ttgctggtga accagcaaac atcgttgctt
     6121 tggttgaaga atacatggat tggttacatc aatctccagt tccaaagttg ttattttggg
     6181 gtacaccagg tgttttgatt ccaccagcag aagctgcaag attggctaag tcattgccaa
     6241 actgtaaggc tgttgatatt ggtccaggtt taaatttgtt acaagaagat aatccagact
     6301 taataggttc cgaaatcgcc agatggttat caactttaga aatcagtggt gctagctaag
     6361 cggccgcgag gagatgaaac aacatattcg aggcaatttt ggcaatatta tagtcattca
     6421 agagaaaaca gtaaatgtat gaaaaatggc aagaacctta tctctatagg cttaataaat
     6481 ctgccttttt ggccaatttt tcaaaacaga tcctatttct tatttataca caatctatat
     6541 ttaacataat taagtacgtg atgacattga agataataat aagtaaagag tttacgtact
     6601 taaattttat atagttttag tatacttgca atgtgtgaat ggatctagta agctgaggta
     6661 cttctctagc gtgaatatga aacgagtaac agggtggtac ataatttacg aacagaactc
     6721 aagttgataa atcctttgcg ttcttaacca tttgtaaaag ttcgtcaaca acattgaaaa
     6781 tattgcttcg agcagatgtt ttatcgtttc tgcaaatttc cagtcctgaa cacctctgtt
     6841 gtcgtaaatg taataatgcc acactggatt cgattctaga agcttggcgt aatcatggtc
     6901 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6961 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7021 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7081 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgc
//
