LOCUS       sequence-201459-        5457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to evaluate the expression output of Saccharomyces arboricola 
            GAL1 promoter with yEGFP used as the reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  AUTHORS   Peng B, Wood RJ, Nielsen LK, Vickers CE
  TITLE     An Expanded Heterologous GAL Promoter Collection for 
            Diauxie-Inducible Expression in Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):748-751. doi: 
            10.1021/acssynbio.7b00355. Epub 2018 Jan 23.
  PUBMED    29301066
REFERENCE   2  (bases 1 to 5457)
  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"; doi: 
            "10.1021/acssynbio.7b00355"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "748-751"
COMMENT     Sequence Label: pILGFP4H
FEATURES             Location/Qualifiers
     source          1..5457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             43..753
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(998..1014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(998..1014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1011..1033)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1022..1038
                     /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(1046..1052,1053..1070,1071..1076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1091..1112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1229..1246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1400..1988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1480..1499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2159..2950,2951..3019))
                     /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     2782..2801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3020..3124)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3192..3210
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3248..3270)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3370..3389
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3583..3605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3597..3614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3598..3614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4728..4761)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 aatactattt atttagggaa atatacaact acaatgggat cctctaaagg tgaagaatta
       61 ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt
      121 tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt
      181 tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt cggttatggt
      241 gttcaatgtt ttgcgagata cccagatcat atgaaacaac atgacttttt caagtctgcc
      301 atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg taactacaag
      361 accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt
      421 attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct
      481 cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt
      541 agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca
      601 attggtgatg gtccagtctt gttaccagac aaccattact tatccactca atctgcctta
      661 tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct
      721 ggtattaccc atggtatgga tgaattgtac aaaagatctt aaactagtag gactatttgc
      781 aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata
      841 tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact
      901 aaactcacaa attagagctt caatttaatt atatcagtta ttacccggga atctcggtcg
      961 taatgattta aatgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1021 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1081 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1141 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1201 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1261 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1321 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1381 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1441 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1501 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1561 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1621 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1681 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1741 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1801 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1861 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1921 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1981 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2041 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2101 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2161 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2221 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2281 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2341 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2401 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2461 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2521 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2581 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2641 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2701 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2761 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2821 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2881 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2941 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3001 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3061 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3121 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     3181 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     3241 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     3301 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     3361 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     3421 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     3481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     3541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     3601 aaacgacggc cagtgaattc atttaaattt ttttgattcg gtaatctccg aacagaagga
     3661 agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga
     3721 aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacct gcaggaaacg
     3781 aagataaaac atgtccacaa aatcatatac cagtagagct gagactcatg caagtccggt
     3841 tgcatcgaaa cttttacgtt taatggatga aaagaagacc aatttgtgtg cttctcttga
     3901 cgttcgttcg actgatgagc tattgaaact tgttgaaacg ttgggtccat acatttgcct
     3961 tttgaaaaca cacgttgata tcttggatga tttcagttat gagggtactg tcgttccatt
     4021 gaaagcattg gcagagaaat acaagttctt gatatttgag gacagaaaat tcgccgatat
     4081 cggtaacaca gtcaaattac aatatacatc gggcgtttac cgtatcgcag aatggtctga
     4141 tatcaccaac gcccacgggg ttactggtgc tggtattgtt gctggcttga aacaaggtgc
     4201 gcaagaggtc accaaagaac caaggggatt attgatgctt gctgaattgt cttccaaggg
     4261 ttctctagca cacggtgaat atactaaggg taccgttgat attgcaaaga gtgataaaga
     4321 tttcgttatt gggttcattg ctcagaacga tatgggagga agagaagaag ggtttgattg
     4381 gctaatcatg accccaggtg taggtttaga cgacaaaggc gatgcattgg gtcagcagta
     4441 cagaaccgtc gacgaagttg taagtggtgg atcagatatc atcattgttg gcagaggact
     4501 tttcgccaag ggtagagatc ctaaggttga aggtgaaaga tacagaaatg ctggatggga
     4561 agcgtaccaa aagagaatca gcgctcccca ttaattatac aggaaactta atagaacaaa
     4621 tcacatattt aatctaatag ccacctgcat tggcacggtg caacactact tcaacttcat
     4681 cttacaaaaa gatcacgtga tctgttgtat tgggatctct agacctaata acttcgtata
     4741 gcatacatta tacgaagtta tattaagggt tgctcgagtt atttttattt ttgtttgatt
     4801 tatccctttc gctatattca aattatagta gaaagtgttg tcaccacatt gcaattcaag
     4861 ctatcctata tttaaacaat ttctattctt acttatatgc catgaaggat ttcaataacc
     4921 cccagtatca ctaccaataa aggaaagcaa tatttaatag cttttactac tatatggtta
     4981 gctgctcact actactttgc aatgttgaag tacggattac ttaccgccga ccggatgaca
     5041 gcactccggc ggaatactct tttccgtgcg tcaccgtctg cgccggtcgt gtccctgaac
     5101 tcagatgtgt ctcgcaccgc catgctccgg acaatgcaaa agtaggaaga ggtgcctgtc
     5161 gtgatggtta tagtgaatgc aaaagacgat tcccctggcc ccgcatactt tgcactcggt
     5221 acaatgcatt gattagcata atgataataa tacgattgag ggggtttcct caccattgta
     5281 atagttgcaa cataaagtgg tggttttcaa tatcttgaca aacatataaa tgcaaatgta
     5341 atattgatta aggtactcag ttttttaatc attccctctt tatagtttta tcacttctag
     5401 ttccacacta ctatgagcta agaaagagat caagagcaaa tttttaatat ctttcgg
//
