LOCUS       sequence-142658-        4593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4593)
  TITLE     monomeric GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4593)
  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            "Unpublished"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: pmGFPS
FEATURES             Location/Qualifiers
     source          1..4593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             12..722
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(39..67)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     662..682
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        863..1050
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1051..1713
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     primer_bind     complement(1977..1993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1977..1993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1990..2012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2001..2017
                     /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(2025..2031,2032..2049,2050..2055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2070..2091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2208..2225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2379..2967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2459..2478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3138..3929,3930..3998))
                     /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     3761..3780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3999..4103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4171..4189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4227..4249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4349..4368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4562..4584
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4576..4593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4577..4593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattcgtgg cagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
       61 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
      121 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
      181 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
      241 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
      301 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
      361 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
      421 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
      481 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
      541 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
      601 accattacct gtccacacaa tctaaacttt cgaaagatcc caacgaaaag agagaccaca
      661 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
      721 aataacccgg gccggggatc cggtgattga ttgagcaagc ttaggagtca ctgccaggta
      781 tcgtttgaac acggcattag tcagggaagt cataacacag tcctttcccg caattttctt
      841 tttctattac tcttggcctc ctctagtaca ctctatattt ttttatgcct cggtaatgat
      901 tttcattttt ttttttccac ctagcggatg actctttttt tttcttagcg attggcatta
      961 tcacataatg aattatacat tatataaagt aatgtgattt cttcgaagaa tatactaaaa
     1021 aatgagcagg caagataaac gaaggcaaag atgacagagc agaaagccct agtaaagcgt
     1081 attacaaatg aaaccaagat tcagattgcg atctctttaa agggtggtcc cctagcgata
     1141 gagcactcga tcttcccaga aaaagaggca gaagcagtag cagaacaggc cacacaatcg
     1201 caagtgatta acgtccacac aggtataggg tttctggacc atatgataca tgctctggcc
     1261 aagcattccg gctggtcgct aatcgttgag tgcattggtg acttacacat agacgaccat
     1321 cacaccactg aagactgcgg gattgctctc ggtcaagctt ttaaagaggc cctaggggcc
     1381 gtgcgtggag taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga
     1441 gcggtggtag atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag
     1501 aaagtaggag atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct
     1561 agcagaatta ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag
     1621 agtgcgttca aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac
     1681 gatgttccct ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca
     1741 tacgatatat atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga
     1801 acagtatgat actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga
     1861 ggcgcgcttt ccttttttct ttttgctttt tctttttttt tctcttgaac tcgaagcttg
     1921 ctcaatcaat caccggatcc gtcgacctgc agccaagcta gcttggcgta atcatggtca
     1981 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2041 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2101 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2161 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2221 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2281 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2341 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2401 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2461 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2521 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2581 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2641 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2701 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2761 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     2821 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2881 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2941 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3001 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3061 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3121 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3181 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3241 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3301 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3361 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3421 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3481 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3541 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3601 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3661 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3721 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3781 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3841 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3901 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3961 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4021 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4081 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4141 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4201 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4261 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4321 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4381 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     4441 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4501 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     4561 tcccagtcac gacgttgtaa aacgacggcc agt
//
