LOCUS       sequence-468715-        3581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-YTK; ConL1/RE cassette; rtTA systems; 
            tetO7-pLEU2m-mNeonGreen-tENO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3581)
  AUTHORS   O'Laughlin R, Tran Q, Lezia A, Ngamkanjanarat W, Emmanuele P, Hao N,
            Hasty J
  TITLE     A Standardized Set of MoClo-Compatible Inducible Promoter Systems 
            for Tunable Gene Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2024 Jan 19;13(1):85-102. doi: 
            10.1021/acssynbio.3c00184. Epub 2023 Dec 11.
  PUBMED    38079574
REFERENCE   2  (bases 1 to 3581)
  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.3c00184"; journalName: "ACS Synth Biol"; date: 
            "2024-01-19- 19"; volume: "13"; issue: "1"; pages: "85-102"
COMMENT     Sequence Label: pRO227
FEATURES             Location/Qualifiers
     source          1..3581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    229..247
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    261..279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    292..310
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    324..342
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    355..373
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    387..405
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    418..436
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(join(1825..2616,2617..2685))
                     /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     2448..2467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2686..2790)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2893..3481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3382..3401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccgcccc tgaattcgca tctagaacgg ggtcatcacg gctcatcatg cgccaaacaa
       61 atgtgtgcaa tacacgctcg gatgactgca tgatgaccgc actgactggg gacagcagat
      121 ccacctaagc ctgtgagaga agcagacacc cgacagatca aggcagttac gtctcaccaa
      181 accagatgtc aacacagcta caacgggtag tccatcgttg taggatactc cctatcagtg
      241 atagagaaat ctatgcggca tccctatcag tgatagagat cggtaacgga gtccctatca
      301 gtgatagaga agtggtgttc agttccctat cagtgataga gaagtttgac actatcccta
      361 tcagtgatag agagaacagc aatgactccc tatcagtgat agagatcaaa tggcgtatcc
      421 ctatcagtga tagagataat taacttgtaa tattctaatc aattgacaat attatttaag
      481 gacctattgt tttttccaat aggtggttag caatcgtctt actttctaac ttttcttacc
      541 ttttacattt cagcaatata tatatatata tttcaaggat ataccattct aagatctatg
      601 gtttcaaaag gggaggagga taatatggcg agtcttcctg ccacgcatga gctgcatatc
      661 ttcggatcta ttaatggtgt cgactttgat atggtaggac aaggaaccgg gaaccccaat
      721 gacgggtatg aagagctaaa tctaaagagt acaaaaggag atttacagtt ttctccttgg
      781 atacttgttc cccacattgg atatggattt catcaatatc ttccttatcc agacggaatg
      841 agcccgttcc aggccgctat ggttgacggc tccgggtatc aagtacaccg taccatgcag
      901 tttgaggatg gggcatccct gaccgtaaat tacaggtaca cctacgaggg ctcacacata
      961 aagggtgagg cgcaggtaaa gggtactgga tttcctgcgg acggcccagt aatgacaaat
     1021 tcacttacag cagcggattg gtgcaggagc aaaaagacct atcccaatga caaaactatc
     1081 ataagtacct ttaaatggtc ctatacgaca ggtaacggga agagatatag gtcaacggca
     1141 aggacaacgt atactttcgc gaagccaatg gcagcaaact atctgaagaa tcaacctatg
     1201 tacgtgttca gaaagaccga gttgaaacat tccaagacag agttaaattt caaagaatgg
     1261 caaaaggctt ttactgacgt aatggggatg gatgaattgt ataagggatc ctaactcgag
     1321 agtgctttta actaagaatt attagtcttt tctgcttatt ttttcatcat agtttagaac
     1381 actttatatt aacgaatagt ttatgaatct atttaggttt aaaaattgat acagttttat
     1441 aagttacttt ttcaaagact cgtgctgtct attgcataat gcactggaag gggaaaaaaa
     1501 aggtgcacac gcgtggcttt ttcttgaatt tgcagtttga aaaatgctga gcatgagacg
     1561 gaaatctgct cgtcagtggt gctcacactg acgaatcatg tacagatcat accgatgact
     1621 gcctggcgac tcacaactaa gcaagacagc cggaaccagc gccggcgaac accactgcat
     1681 atatggcata tcacaacagt ccaactagtg cactgcagta cagcggccgc gattatcaaa
     1741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1861 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1921 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc cacgctcacc
     1981 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2041 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2101 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2161 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2221 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2281 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2341 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2401 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2461 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2521 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2581 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2641 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2701 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2761 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgtcat
     2821 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2881 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2941 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3001 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3061 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3121 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3181 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3241 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3301 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3361 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3421 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3481 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3541 gttctttcct gcgttatccc ctgattctgt ggataaccgt g
//
