LOCUS       sequence-124239-        2320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The second of the two RNA lassos required for the RNA chain 
            construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2320)
  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 2320)
  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: Lasso 2
FEATURES             Location/Qualifiers
     source          1..2320
                     /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      219..266
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      282..340
                     /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(364..381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(535..1123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(615..634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1306..1400)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1421..2080)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1995..2014
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2242..2260
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2264..2307)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 cgcttaatac gactcactat agggaggctg tcctcgtccg tatgacgaga gaagcctacc
       61 agagaaacac acgacgtaag tcgtggtaca ttacctggta caagccttgt cccttgaaga
      121 gaacaactaa caactaacaa caacaatcaa ctattaaacg gtagacccaa acatacaaca
      181 acaatcaaca atcaaccaag gctgtcctcg tcgatatcct agcataaccc cttggggcct
      241 ctaaacgggt cttgaggggt tttttgagcc tcttcctgca gtccggcaaa aaaacggcaa
      301 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc
      361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      541 caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1261 ctaaagtata tatgagtaaa cttggtctga cagctcgagg cttggattct caccaataaa
     1321 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     1381 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
     1441 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     1501 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     1561 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     1621 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     1681 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     1741 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     1801 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     1861 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     1921 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     1981 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     2041 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     2101 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     2161 tggaacctct tacgtgcccg atcaactcga gtgccacctg acgtctaaga aaccattatt
     2221 atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa
     2281 tccttagctt tcgctaagga tgatttctgg aattcgaaga
//
