LOCUS       sequence-376671-        3796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control, empty integrating inducible casette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3796)
  AUTHORS   Lopez SC, Crawford KD, Lear SK, Bhattarai-Kline S, Shipman SL
  TITLE     Precise genome editing across kingdoms of life using retron-derived 
            DNA.
  JOURNAL   Nat Chem Biol. 2022 Feb;18(2):199-206. doi: 
            10.1038/s41589-021-00927-y. Epub 2021 Dec 23.
  PUBMED    34949838
REFERENCE   2  (bases 1 to 3796)
  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: 
            "10.1038/s41589-021-00927-y"; journalName: "Nat Chem Biol"; date: 
            "2022-02"; volume: "18"; issue: "2"; pages: "199-206"
COMMENT     Sequence Label: pSCL.004
FEATURES             Location/Qualifiers
     source          1..3796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(26..614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(106..125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(785..1576,1577..1645))
                     /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     1408..1427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1646..1750)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1818..1836
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1874..1896)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1996..2015
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2162..2348
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2349..3011
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(3271..3726)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3408..3429)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3620..3639
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
       61 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      121 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      181 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      241 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      301 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      361 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      421 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      481 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      541 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      601 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      661 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      721 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      781 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
      841 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
      901 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
      961 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1021 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1081 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1141 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1201 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1261 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1321 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1381 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1441 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1501 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1561 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1621 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1681 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1741 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     1801 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     1861 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     1921 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     1981 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaattcccgt
     2041 tttaagagct tggtgagcgc taggagtcac tgccaggtat cgtttgaaca cggcattagt
     2101 cagggaagtc ataacacagt cctttcccgc aattttcttt ttctattact cttggcctcc
     2161 tctagtacac tctatatttt tttatgcctc ggtaatgatt ttcatttttt ttttccacct
     2221 agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa ttatacatta
     2281 tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca agataaacga
     2341 aggcaaagat gacagagcag aaagccctag taaagcgtat tacaaatgaa accaagattc
     2401 agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc ttcccagaaa
     2461 aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac gtccacacag
     2521 gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc tggtcgctaa
     2581 tcgttgagtg cattggtgac ttacacatag acgaccatca caccactgaa gactgcggga
     2641 ttgctctcgg tcaagctttt aaagaggccc taggggccgt gcgtggagta aaaaggtttg
     2701 gatcaggatt tgcgcctttg gatgaggcac tttccagagc ggtggtagat ctttcgaaca
     2761 ggccgtacgc agttgtcgaa cttggtttgc aaagggagaa agtaggagat ctctcttgcg
     2821 agatgatccc gcattttctt gaaagctttg cagaggctag cagaattacc ctccacgttg
     2881 attgtctgcg aggcaagaat gatcatcacc gtagtgagag tgcgttcaag gctcttgcgg
     2941 ttgccataag agaagccacc tcgcccaatg gtaccaacga tgttccctcc accaaaggtg
     3001 ttcttatgta gtgacaccga ttatttaaag ctgcagcata cgatatatat acatgtgtat
     3061 atatgtatac ctatgaatgt cagtaagtat gtatacgaac agtatgatac tgaagatgac
     3121 aaggtaatgc atcattctat acgtgtcatt ctgaacgagg cgcgctttcc ttttttcttt
     3181 ttgctttttc tttttttttc tcttgaactc gacggatcta tgcggtgtga aataccgcac
     3241 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     3301 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3361 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3421 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3481 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3541 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3601 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3661 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3721 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     3781 ttcgctatta cgccag
//
