LOCUS       sequence-492077-        7261 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  Insert an N-terminal msGFP2 tag at TMN1 and replace native 
            presequence with OST1 presequence by popin-popout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7261)
  TITLE     Glick lab plasmids 2025
REFERENCE   2  (bases 1 to 7261)
  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-pOst1-msGFP2-Tmn1
FEATURES             Location/Qualifiers
     source          1..7261
                     /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             1084..1788
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="DSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITGGSGGS"
     primer_bind     complement(3758..3775)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3758..3774)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3767..3789)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3983..4002)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4102..4124
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4242..5045)
                     /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(5046..5262)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5269..5287)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5355..5459
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5460..5528,5529..6320)
                     /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(5678..5697)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6491..7079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6980..6999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7233..7250
                     /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 ccaagcttgc
      241 atgcctgcag gtcgactcta gatcatccca gcctccatct tcatcggcaa gcagtgaatc
      301 taaatcaaaa tcgaagaaga agaaaatagg tcgtggtttt gtcgttttaa ttggtcttgc
      361 actagcatta gggacagttt ctgtccttgg tattgcaggc gtcattcttg cctacgtgtt
      421 taaggatcca gagggtgatt ataagcccat taaaccgcga attgatgaga atgaaatgct
      481 tgataccgtc ccacccgaaa aacttatgaa gtttgtctaa tcggggttgt attatataat
      541 atataactac ttaaaataat gaacacttaa tataataaac aaaaataaaa taggaaaatc
      601 ctaagttgat caaaacttaa taaaaattac tacttacact ggaatgttag tagtaaattc
      661 aacacgtatt ggaagtcact tggaatgtta ccaatactgc ctgagaatga gtgattttaa
      721 cttataatac aaaagtccgg gttacagccc tttaaatttt tttcacggca atttttcttt
      781 acttggtgaa gccatgaaaa atgtcatcgg ttcgaagaaa taggcctaat gataaacaaa
      841 aaggtaattc aaggtttttg aactgctttg ttttttttat ttgtatcttt ttttggtacg
      901 ttagttaagt taatttttgg ctccttgtcc agcttttcct tcatctgcta ccgttatatt
      961 ttcagatatt aatttagtag cagagagcaa gtaccgaaag gaaaacaaaa cagaatgagg
     1021 caggtttggt tctcttggat tgtgggattg ttcctatgtt ttttcaacgt gtcttctgct
     1081 gccgatagca ctgagagctt gttcacaggt gttgttccta tcttggtcga attagacggt
     1141 gacgtaaacg gtcataagtt cagtgttaga ggtgaaggtg aaggtgacgc tactaacggt
     1201 aaattgacat tgaagtttat ttgtactaca ggtaaattgc cagttccttg gccaactttg
     1261 gttactacat tgacatacgg tgttcaatgt ttttcaagat acccagatca tatgaagcaa
     1321 catgatttct ttaagtctgc tatgccagaa ggttacgttc aagaaagaac tatcactttt
     1381 aaagatgatg gtacttacaa gacaagagca gaggttaagt tcgaaggtga cactttggtt
     1441 aacagaatcg aattgaaggg tatcgatttt aaagaagatg gtaatatttt gggtcataaa
     1501 ttggaataca actacaactc acataacgtt tacattacag ctgataaaca aaagaatggt
     1561 attaaagcaa acttcaagat cagacataat gttgaagatg gttctgttca attagcagat
     1621 cattaccaac aaaacactcc aattggtgac ggtccagttt tgttaccaga taaccattac
     1681 ttgtctacac aatcaaaatt gtctaaagat ccaaacgaaa agagagatca tatggttttg
     1741 ttagaatttg ttactgctgc tggtatcaca ggtggcagtg gcggttctgg aggctcagga
     1801 gggagcttct atttaccagg tgtagcgcct accacttaca gggaaaatga caacatccca
     1861 ttgctcgtta atcatttaac tccatcaatg aattatcaac acaaagatga ggacggtaat
     1921 aacgtttctg gcgataaaga aaatttcttg tactcatacg actactatta caatagattc
     1981 catttttgtc agcctgaaaa agtcgaaaag caacctgaat ccttgggatc cgttatattt
     2041 ggtgatcgaa tttataattc accattccaa ttgaatatgt tacaagaaaa agagtgtgaa
     2101 tccctgtgta agaccgtaat tccgggagat gatgctaaat tcatcaataa attgataaaa
     2161 aatggcttct tccaaaactg gcttattgat ggattaccgg ctgcaagaga ggtttatgat
     2221 ggcagaacta agactagttt ttatggtgct ggcttcaacc ttggttttgt acaggtaact
     2281 caaggaaccg acattgaggc tactccaaaa ggggcagaaa ccaccgataa ggatgtggaa
     2341 ttggaaacac gtaatgatcg taacatggtg aaaacttacg aattgcctta ctttgccaac
     2401 catttcgata ttatgattga atatcacgat cgtggtgaag gtaactatcg cgtagttggt
     2461 gttatcgtcg aaccagtttc cataaaacgt tccagtccag gtacatgtga aacgactggc
     2521 agcccattga tgctagacga agaaaatgat aatgaagttt attttactta ttccgttaaa
     2581 ttcaatgaat cagcgacttc ttgggccacc aggtgggaca aatatttaca cgtatatgat
     2641 ccttctattc aatggttttc tttgataaat ttctccttag ttgttgtctt attgtcatct
     2701 gttgttatcc attcgctatt gcgcgcactg aaaagtgatt tcgcacgtta caacgagtta
     2761 aatttggacg acgattttca agaagattca ggttggaaat tgaatcatgg tgatgttttt
     2821 cgttcgccaa gtcaatcgct aaccctttca attttggttg gttcaggagt tcaattattt
     2881 ttgatggtaa cttgcagtat cttcttcgct gcattgggtt tcctatcacc tagttctaga
     2941 ggttccttag ccactgttat gtttatctta tatgcattat tcgggtttgt cggctcttat
     3001 acatctatgg gcatctacaa atttttcaat gggccatact ggaaggcaaa cttgatattg
     3061 actccacttc tagttcctgg tgctattctg ctaataatta ttgcgttgaa ctttttttta
     3121 atgtttgttc attcttctgg tgtcatacca gcaagtactc tgttttttat ggtttttcta
     3181 tggtttttgt tctcaattcc attatcattt gctggttcat tgattgccag gaaaagatgt
     3241 cactgggatg aacatccaac caagacaaac caaatcgcaa gacaaatacc cttccagcct
     3301 tggtatttaa agactatacc agctactttg attgctggta tctttccatt tggctccatc
     3361 gccgttgagt tgtatttcat atacacaagt ttatggttta ataagatttt ttacatgttt
     3421 ggtttcctct ttttctcttt cctcctatta actttgacaa gttcactcgt cactattttg
     3481 atcacttatc attccttatg cctggagaac tggaagtggc aatggagagg attcataatt
     3541 ggaggtgccg gttgtgcttt gtatgtgttt attcattcta ttttattcac caaatttaag
     3601 ttgggtggat tcacgactat tgtcttatac gtgggatact cctcagtcat ttcattactc
     3661 tgctgtttgg taacggggtc aattggcttt atcagcagta tgctctttgt tagaaaaatc
     3721 tactcatcca ttaaagttga ctagcgagct cgaattcact ggccgtcgtt ttacaacgtc
     3781 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     3841 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3901 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3961 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4021 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4081 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4141 cgaaacgcgc gagacgaaag ggcgggtaat aactgatata attaaattga agctctaatt
     4201 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     4261 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     4321 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     4381 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     4441 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     4501 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     4561 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     4621 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     4681 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     4741 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     4801 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     4861 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     4921 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     4981 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     5041 gacatgattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt taagaatact
     5101 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     5161 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     5221 cgaaatcaaa aaaaagaata aaaaaaaaaa tgatgaattg aaaagctggc ctcgtgatac
     5281 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5341 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5401 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5461 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5521 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5581 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5641 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5701 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5761 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5821 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5881 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5941 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6001 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6061 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6121 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6181 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6241 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6301 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6361 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6421 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6481 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6541 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6601 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6661 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6721 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6781 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6841 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6901 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6961 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7021 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7081 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7141 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7201 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7261 a
//
