LOCUS       sequence-326015-        2814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2814)
  AUTHORS   Fulk EM, Huh D, Atkinson JT, Lie M, Masiello CA, Silberg JJ
  TITLE     A Split Methyl Halide Transferase AND Gate That Reports by 
            Synthesizing an Indicator Gas.
  JOURNAL   ACS Synth Biol. 2020 Nov 20;9(11):3104-3113. doi: 
            10.1021/acssynbio.0c00355. Epub 2020 Oct 26.
  PUBMED    33104325
REFERENCE   2  (bases 1 to 2814)
  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.0c00355"; journalName: "ACS Synth Biol"; date: 
            "2020-11-20- 20"; volume: "9"; issue: "11"; pages: "3104-3113"
COMMENT     Sequence Label: pEMF038
FEATURES             Location/Qualifiers
     source          1..2814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(37..696)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     611..630
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     865..883
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(904..922,923..928,929..945,946..951,952..977)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    904..922
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    929..947
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1108..1142
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1177..1803
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1253..1272)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      1809..1895
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2060..2648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2140..2159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(join(2736..2814,1..16))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc
       61 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
      121 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
      181 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
      241 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
      301 ggccaggttt tcaccataac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
      361 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
      421 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa
      481 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
      541 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
      601 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
      661 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
      721 tctcgataac tcaaaaaata cgaccggtag tgatcttatt tcattatggt gaaagttgga
      781 acctcttacg ggccggccaa cgtctcattt tcgccagata tcgacgtcta agaaaccatt
      841 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc
      901 gagtccctat cagtgataga gattgacatc cctatcagtg atagagatac tgagcacatc
      961 agcaggacgc actgaccgat aggacccata caaaggaagt tttgtaataa tctaactaaa
     1021 aaggcctccc aaatcggggg gcctttttta ttgataacaa aaggtacccg tggatcctct
     1081 agagtcgacc tgcaggcatg caagcttttt acggctagct cagtcctagg tatagtgcta
     1141 gcccagccag agaaacactc tttaacaggg ggtagtatga tgtctcgttt agataaaagt
     1201 aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc
     1261 cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat
     1321 aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt
     1381 tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga
     1441 tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa
     1501 aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta
     1561 gagaatgcat tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa
     1621 gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg
     1681 ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta
     1741 ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct
     1801 taaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1861 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc tagggcgttc
     1921 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1981 gggataacgc atgcaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2041 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgtc gcttaccgga tacctgtccg
     2221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2461 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     2521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2581 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2641 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2701 cacgttaagg gattttggtc atgactagtg cttggattct caccaataaa aaacgcccgg
     2761 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tgga
//
