LOCUS       sequence-371335-        5184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration cassette to create C-terminal fusions for fluorescent 
            tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5184)
  AUTHORS   Baek I, Le SN, Jeon J, Chun Y, Reed C, Buratowski S
  TITLE     A set of Saccharomyces cerevisiae integration vectors for 
            fluorescent dye labeling of proteins.
  JOURNAL   G3 (Bethesda). 2022 Aug 9. pii: 6659097. doi: 
            10.1093/g3journal/jkac201.
  PUBMED    35944214
REFERENCE   2  (bases 1 to 5184)
  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: "G3 
            (Bethesda). 2022 Aug 9. pii: 6659097. doi: 
            10.1093/g3journal/jkac201."
COMMENT     Sequence Label: pBS-SKII-3XHA-HALO-NAT
FEATURES             Location/Qualifiers
     source          1..5184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5..94
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             125..1015
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     gene            1073..2192
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1073..1416
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1417..1989
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1995..2192
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2261..2280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2262..2280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2290..2307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2290..2306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2299..2321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2448..2903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2535..2554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2745..2766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2929..3033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3034..3102,3103..3894)
                     /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(3252..3271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4065..4653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4554..4573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4807..4824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4941..4962
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4977..4982,4983..5000,5001..5007)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5015..5031
                     /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     5020..5042
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5039..5055
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5039..5055
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5074..5094
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5076..5094
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     5123..5139
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5124..5140
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cttttaccca tacgatgttc ctgactatgc gggctatccc tatgacgtcc cggactatgc
       61 aggatcctat ccatatgacg ttccagatta cgctgctcag tgcggttcag gtggatctgg
      121 ttctatggca gaaatcggta ctggctttcc attcgacccg cattatgtgg aagtcctggg
      181 cgagagaatg cattacgttg acgtgggtcc gagagatgga actccggtcc tttttctgca
      241 cgggaatcct acaagctctt atgtttggcg caatatcatc cctcatgtag ctccgacgca
      301 tcgctgtatt gcgccggacc tgattggtat gggaaaatct gataaaccag acctgggtta
      361 ctttttcgat gatcatgtgc gtttcatgga tgccttcatt gaggcattag ggcttgaaga
      421 agtcgtcttg gtgattcatg attggggctc agctctggga tttcactggg ctaaaagaaa
      481 tcctgaacgc gtaaaaggca tcgcgtttat ggagttcatt cgtccaattc cgacttggga
      541 tgaatggcct gagttcgcga gagaaacatt tcaagcattt cgcacgaccg atgtaggccg
      601 gaagttaatc atcgatcaga atgtctttat cgaagggaca ttgccgatgg gagtcgttcg
      661 tccgttaaca gaagtcgaaa tggatcacta tagagaacct tttcttaatc ctgtggacag
      721 agagccgctg tggcggtttc cgaacgaact gccgattgca ggcgagcctg ctaacattgt
      781 agcgctggtt gaagagtata tggattggct tcatcagtct ccagttccga agttattgtt
      841 ttggggtacg cctggcgtgc ttattccacc ggccgaagcg gcacgtttgg caaaaagcct
      901 gccaaattgc aaagccgttg acattggccc tggacttaac ttgcttcaag aggataaccc
      961 ggacttaatc gggagcgaaa ttgcccggtg gctttctacc ttagaaatca gcggctaata
     1021 tataactgtc tagaaataaa gagtatcatc tttcaaaggt cacccggcca gcgacatgga
     1081 ggcccagaat accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg
     1141 tcgcccgtac atttagccca tacatcccca tgtataatca tttgcatcca tacattttga
     1201 tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa
     1261 cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa
     1321 aggttaggat ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg
     1381 atacagttct cacatcacat ccgaacataa acaaccatgg gtaccactct tgacgacacg
     1441 gcttaccggt accgcaccag tgtcccgggg gacgccgagg ccatcgaggc actggatggg
     1501 tccttcacca ccgacaccgt cttccgcgtc accgccaccg gggacggctt caccctgcgg
     1561 gaggtgccgg tggacccgcc cctgaccaag gtgttccccg acgacgaatc ggacgacgaa
     1621 tcggacgacg gggaggacgg cgacccggac tcccggacgt tcgtcgcgta cggggacgac
     1681 ggcgacctgg cgggcttcgt ggtcgtctcg tactccggct ggaaccgccg gctgaccgtc
     1741 gaggacatcg aggtcgcccc ggagcaccgg gggcacgggg tcgggcgcgc gttgatgggg
     1801 ctcgcgacgg agttcgcccg cgagcggggc gccgggcacc tctggctgga ggtcaccaac
     1861 gtcaacgcac cggcgatcca cgcgtaccgg cggatggggt tcaccctctg cggcctggac
     1921 accgccctgt acgacggcac cgcctcggac ggcgagcagg cgctctacat gagcatgccc
     1981 tgcccctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat
     2041 agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt
     2101 ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg
     2161 tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccact
     2221 agttctagag cggccgccac cgcggtggag ctccaattcg ccctatagtg agtcgtatta
     2281 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2341 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2401 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     2461 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2521 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2581 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2641 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2701 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2761 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2821 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2881 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     2941 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3001 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3061 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3121 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3181 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3241 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3301 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3361 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3421 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3481 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3541 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3601 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3661 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3721 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3781 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3841 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3901 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3961 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4021 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4081 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4141 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4201 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4261 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4321 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4381 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4441 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4501 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4561 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4621 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4681 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4741 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4801 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4861 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4921 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     4981 cactttatgc tcccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     5041 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     5101 aagctgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgggtcg
     5161 acggatcccc gggttaatta acat
//
