LOCUS       sequence-125474-        3221 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 3221)
  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 3221)
  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: P4U2C8T2-v1-1 (Pl-tetO1-deCFP) (135p)
FEATURES             Location/Qualifiers
     source          1..3221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    25..43
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    50..68
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             92..114
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(151..172)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(412..431)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     759..780
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(994..1011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1165..1753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1245..1264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1924..2715,2716..2784))
                     /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     2547..2566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2785..2889)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2957..2975
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcattgctgt tccgctgggc atgctcccta tcagtgatag agattgacat ccctatcagt
       61 gatagagata ctgagcacag caagcaataa ttttgtttaa ctttaagaag gagatatacc
      121 atggagcttt tcactggcgt tgttcccatc ctggtcgagc tggacggcga cgtaaacggc
      181 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      241 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      301 acctggggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      361 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      421 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      481 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      541 tacatcagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
      601 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      661 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      721 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      781 accgccgccg ggatctaatg aagcatctgg tgaataactc gagcaaagcc cgccgaaagg
      841 cgggcttttc tgtgtcgacc gatgcccttg agagtcgggc attgtcttcg ctccttccgg
      901 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tgttctttat catgcaactc
      961 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1021 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1081 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1561 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1801 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1861 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1921 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1981 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2041 ccccagtgct gcaatgatac cgcgtgaccc acgctcaccg gctccagatt tatcagcaat
     2101 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2161 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2221 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2281 attcagctcc ggttcccaac gatcaaggcg agttgcatga tcccccatgt tgtgcaaaaa
     2341 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2401 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2461 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2521 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     2581 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2641 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2701 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2761 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2821 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2881 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     2941 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtgttcaag aattctggcg
     3001 aatcctctga ccagccagaa aacgaccttt ctgtggtgaa accggatgct gcaattcaga
     3061 gcgccagcaa gtgggggaca gcagatgacc tgaccgccgc agagtggatg tttgacatgg
     3121 tgatgactat cgcaccatca gccagaaaac cgaattttgc tgggtgggct aacgatatcc
     3181 gcctgatgcg tgaacgtgac ggacgtaacg aagacaacca c
//
