LOCUS       sequence-110249-        2789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes AmpR-ColE1 as a Type 8a part to be used in the Dueber YTK 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2789)
  AUTHORS   Lee ME, DeLoache WC, Cervantes B, Dueber JE
  TITLE     A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
  JOURNAL   ACS Synth Biol. 2015 Apr 14.
  PUBMED    25871405
REFERENCE   2  (bases 1 to 2789)
  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: "ACS Synth 
            Biol. 2015 Apr 14."
COMMENT     Sequence Label: pYTK089
FEATURES             Location/Qualifiers
     source          1..2789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(116..907,908..976))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     739..758
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(977..1081)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1184..1772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1673..1692
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1893..1911
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1918..1936
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1954..1965
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1972..2649
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2667..2738
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2754..2781
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tcggtctcac cgagcggccg cgattatcaa aaaggatctt cacctagatc cttttaaatt
       61 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
      121 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      181 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      241 ctgcaatgat accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc
      301 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      361 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      421 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      481 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      541 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      601 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      661 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      721 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      781 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      841 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
      901 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      961 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1021 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1081 gcacatttcc ccgaaaagtg ccacctgtca tgaccaaaat cccttaacgt gagttttcgt
     1141 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     1201 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1261 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1321 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1381 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     1441 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     1501 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     1561 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     1621 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     1681 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     1741 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     1801 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     1861 tggataaccg tgcggccgcc aattgagacc agtccctatc agtgatagag attgacatcc
     1921 ctatcagtga tagagatact gagcacggat ctgaaagagg agaaaggatc tatggcgagt
     1981 agcgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac
     2041 ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg tactcagacc
     2101 gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat cctgtccccg
     2161 cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc ggactacctg
     2221 aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga agacggtggt
     2281 gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta caaagttaaa
     2341 ctgcgtggta ctaacttccc gtccgacggt ccggttatgc agaaaaaaac catgggttgg
     2401 gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg
     2461 cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac ctacatggct
     2521 aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga catcacctcc
     2581 cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc
     2641 ggtgcttaat aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2701 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     2761 cttcgggtgg gcctttctgc gtttataag
//
