LOCUS       sequence-125477-        3167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1 Part of 4-part switch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3167)
  AUTHORS   Sun ZZ, Yeung E, Hayes CA, Noireaux V, Murray RM
  TITLE     Linear DNA for rapid prototyping of synthetic biological circuits in
            an Escherichia coli based TX-TL cell-free system.
  JOURNAL   ACS Synth Biol. 2014 Jun 20;3(6):387-97. doi: 10.1021/sb400131a. 
            Epub 2013 Dec 4.
  PUBMED    24303785
REFERENCE   2  (bases 1 to 3167)
  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/sb400131a";
            journalName: "ACS Synth Biol"; date: "2014-06-20- 20"; volume: "3"; 
            issue: "6"; pages: "387-97"
COMMENT     Sequence Label: P3U2C5T2-v1-2 (Pl-lacO1-tetR) (136p)
FEATURES             Location/Qualifiers
     source          1..3167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(121..138)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(292..880)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(372..391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1051..1842,1843..1911))
                     /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     1674..1693
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1912..2016)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2084..2102
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(2427..3050)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     2958..2977
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     RBS             3057..3079
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3105..3121
                     /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)."
ORIGIN
        1 atgctatgtt gtcttcgctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
       61 atgactgtgt tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
      121 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      181 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      241 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      301 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      361 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      421 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      481 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      541 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      601 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      661 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      721 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      781 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      841 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      901 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      961 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1021 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1081 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1141 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gtgacccacg
     1201 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1261 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1321 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1381 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1441 tgcatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1501 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1561 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1621 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     1681 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1741 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1801 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1861 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1921 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1981 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2041 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     2101 gccctttcgt gttcaagaat tctggcgaat cctctgacca gccagaaaac gacctttctg
     2161 tggtgaaacc ggatgctgca attcagagcg ccagcaagtg ggggacagca gatgacctga
     2221 ccgccgcaga gtggatgttt gacatggtga tgactatcgc accatcagcc agaaaaccga
     2281 attttgctgg gtgggctaac gatatccgcc tgatgcgtga acgtgacgga cgtaacgaag
     2341 acaaggcacg actctcaagg gcatcggtcg acacagaaaa gcccgccttt cggcgggctt
     2401 tgctcgagtt attcaccaga gcttcattaa gacccacttt cacatttaag ttgtttttct
     2461 aatccgcata tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca
     2521 aataattcga tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt
     2581 tcttctttag cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc
     2641 acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct
     2701 aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt
     2761 gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa
     2821 tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca
     2881 atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc
     2941 tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta
     3001 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat ggtatatctc
     3061 cttcttaaag ttaaacaaaa ttattgcttg ctgtgctcag tatcttgtta tccgctcaca
     3121 atgtcaatgt tatccgctca catttatgca tgcccagcgg aacagca
//
