LOCUS       sequence-124245-        2268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct targeting the PIE-target sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2268)
  AUTHORS   Hassall J, MacDonald P, Cordero T, Rostain W, Jaramillo A
  TITLE     Design and Characterization of Topological Small RNAs.
  JOURNAL   Methods Mol Biol. 2015;1316:149-67. doi: 
            10.1007/978-1-4939-2730-2_13.
  PUBMED    25967060
REFERENCE   2  (bases 1 to 2268)
  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: "Methods Mol
            Biol."; date: "2015"; volume: "1316:149-67. doi"; pages: " 
            10.1007/978-1-4939-2730-2_13"
COMMENT     Sequence Label: AL562-2
FEATURES             Location/Qualifiers
     source          1..2268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..24
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5..23
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      167..214
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      230..288
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(312..329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(483..1071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(563..582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1254..1348)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1369..2028)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1943..1962
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2190..2208
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2212..2255)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 cgcttaatac gactcactat agggaggctg tcctcgtccg tatgacgaga gaagcctacc
       61 agagaaacac acgacgtaag tcgtggtaca ttacctggta catattaaac ggtagaccca
      121 aacaacaaca agaacaacaa caaccaaggc tgtcctcgtc tgcgcactag cataacccct
      181 tggggcctct aaacgggtct tgaggggttt tttgagcctc ttcctgcagt ccggcaaaaa
      241 aacggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt
      301 atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      481 tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      901 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgaggct tggattctca
     1261 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     1321 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     1381 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     1441 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     1501 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     1561 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     1621 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     1681 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     1741 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     1801 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     1861 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     1921 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     1981 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     2041 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     2101 gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac gtctaagaaa
     2161 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat
     2221 aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgaaga
//
