LOCUS       sequence-125476-        3626 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 3626)
  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 3626)
  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: P4U2C6T2-v1-2 (Pl-tetO1-lacI) (137p)
FEATURES             Location/Qualifiers
     source          1..3626
                     /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(2428..3510)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3472..3491
                     /label=LacI-R
                     /note="LacI, reverse primer"
     RBS             3517..3539
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3563..3581
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3588..3606
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
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 tgcttcatca ctgcccgctt tccagtcggg aaacctgtcg
     2461 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2521 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     2581 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     2641 tttgatggtg gttaacggcg ggatataaca tgagctgtcc tcggtatcgt cgtatcccac
     2701 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     2761 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     2821 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     2881 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     2941 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     3001 gcccagtcgc gtaccgtcct catgggagaa aataatactg ttgatgggtg tctggtcaga
     3061 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     3121 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     3181 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     3241 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     3301 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     3361 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     3421 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     3481 tgcgacatcg tataacgtta ctggtttcat ggtatatctc cttcttaaag ttaaacaaaa
     3541 ttattgcttg ctgtgctcag tatctctatc actgataggg atgtcaatct ctatcactga
     3601 tagggagcat gcccagcgga acagca
//
