LOCUS       sequence-126616-        4539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Citrine reporter for transfer fxns.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4539)
  AUTHORS   Niederholtmeyer H, Sun ZZ, Hori Y, Yeung E, Verpoorte A, Murray RM, 
            Maerkl SJ
  TITLE     Rapid cell-free forward engineering of novel genetic ring 
            oscillators.
  JOURNAL   Elife. 2015 Oct 2;4. doi: 10.7554/eLife.09771.
  PUBMED    26430766
REFERENCE   2  (bases 1 to 4539)
  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: "Elife."; 
            date: "2015-10-2"; volume: "4. doi"; pages: " 10.7554/eLife.09771"
COMMENT     Sequence Label: pLacI-BCD7-Citrine P42U18C68T11 v40-1 (536)
FEATURES             Location/Qualifiers
     source          1..4539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    128..144
                     /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)."
     CDS             249..965
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSRLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1061..1132
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1148..1175
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1706..2656)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(3418..3512)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3543..4334,4335..4403))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4166..4185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgaa gacaaccacg cattgctgtt cataaatgtg agcggataac
      121 attgacattg tgagcggata acaagatact gagcacagca agcgggccca agttcactta
      181 aaaaggagat caacaatgaa agcaattttc gtactgaaac atcttaatca tgctggggag
      241 ggtttctaat gtctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat
      301 tagatggtga tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta
      361 cttacggtaa attgacctta aaatttattt gtactactgg taaattgcca gttccatggc
      421 caaccttagt cactacttta ggttatggtt tgatgtgttt tgctagatac ccagatcata
      481 tgaaacaaca tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta
      541 tttttttcaa agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata
      601 ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag
      661 gtcacaaatt ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa
      721 agaatggtat caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat
      781 tagctgacca ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca
      841 accattactt atcctatcaa tctagattat ccaaagatcc aaacgaaaag agagatcaca
      901 tggtcttgtt agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca
      961 aatgatgaac tcaaagcccg ccgaaaggcg ggcttttctg tccttgagag tcgggcattg
     1021 tcttctaagg atccaaactc gagtaaggat ctccaggcat caaataaaac gaaaggctca
     1081 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     1141 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acctagggta cgggttttgc
     1201 tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt
     1261 cagccggttt gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac
     1321 gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg
     1381 tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc
     1441 atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc
     1501 atctgttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa
     1561 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc
     1621 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
     1681 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     1741 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
     1801 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     1861 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
     1921 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     1981 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     2041 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     2101 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     2161 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     2221 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     2281 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     2341 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     2401 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     2461 cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc
     2521 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     2581 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     2641 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     2701 gatgggctag tcaatgataa ttactagtcc ttttcccggg tgatctgggt atctgtaaat
     2761 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     2821 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     2881 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     2941 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
     3001 aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt
     3061 ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg
     3121 ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat
     3181 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
     3241 gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat
     3301 tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc
     3361 cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt
     3421 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     3481 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc
     3541 gcttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3601 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3661 cccagtgctg caatgatacc gcgtgaccca cgctcaccgg ctccagattt atcagcaata
     3721 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3781 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3841 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3901 ttcagctccg gttcccaacg atcaaggcga gttgcatgat cccccatgtt gtgcaaaaaa
     3961 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4021 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4081 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4141 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     4201 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4261 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4321 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4381 acacggaaat gttgaatact catgcgaaac gatcctcatc ctgtctcttg atcagatcat
     4441 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
     4501 ttcccaacct taccagaggg cgccccagct ggcaattcc
//
