LOCUS       sequence-435422-        3701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Theophylline_induced self_splicing intron_based riboswitch for the 
            moderate expression of Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3701)
  AUTHORS   Xu J, Sun Y, Wu J, Yang S, Yang L
  TITLE     Chromosome recombination and modification by LoxP-mediated evolution
            in Vibrio natriegens using CRISPR-associated transposases.
  JOURNAL   Biotechnol Bioeng. 2024 Mar;121(3):1163-1172. doi: 
            10.1002/bit.28639. Epub 2023 Dec 22.
  PUBMED    38131162
REFERENCE   2  (bases 1 to 3701)
  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: "10.1002/bit.28639";
            journalName: "Biotechnol Bioeng"; date: "2024-03"; volume: "121"; 
            issue: "3"; pages: "1163-1172"
COMMENT     Sequence Label: pSIBR4-Cre
FEATURES             Location/Qualifiers
     source          1..3701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..26
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        join(33..38,39..56,57..63)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     complement(1522..1539)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1692..1778
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1880..2539)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2454..2473
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2540..2642)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2919..2936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3053..3598)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3133..3152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaagcggat aacaatttca cacaggaggc cgtttacact ttatgcttcc ggctcgtata
       61 atgtgtgggg aaaccacaac ggtttccctc tagaaataat tttgtttaac tataagaagg
      121 agatatacat atgtcctctg gttaattgag gcctgagtat aaggtgactt atacttgtaa
      181 tctatctaaa cggggaacct ctctagtaga caatcccgtg ctaaattgat accagcatcg
      241 tcttgatgcc cttggcagca taaatgccta acgactatcc ctttggggag tagggtcaag
      301 tgactcgaaa cgatagacaa cttgctttaa caagttggag atatagtctg ctctgcatgg
      361 tgacatgcag ctggatataa ttccggggta agattaacga ccttatctga acataatgct
      421 aagcaatctg ctgaccgttc accaaaatct gccggcgctg ccggttgatg cgaccagcga
      481 cgaagtgcgt aaaaatctga tggacatgtt ccgtgaccgt caggcgttta gcgagcacac
      541 ctggaagatg ctgctgagcg tgtgccgtag ctgggcggcg tggtgcaagc tgaacaaccg
      601 taaatggttt ccggcggagc cggaagatgt gcgtgattac ctgctgtatc tgcaggcgcg
      661 tggcctggcg gttaaaacca ttcagcaaca cctgggtcaa ctgaacatgc tgcaccgtcg
      721 tagcggcctg ccgcgtccga gcgacagcaa cgcggtgagc ctggttatgc gtcgtatccg
      781 taaggagaac gtggatgcgg gtgaacgtgc gaaacaggcg ctggcgttcg agcgtaccga
      841 ctttgatcaa gttcgtagcc tgatggaaaa cagcgaccgt tgccaagata ttcgtaacct
      901 ggcgtttctg ggcatcgcgt acaacaccct gctgcgtatc gcggagattg cgcgtatccg
      961 tgttaaggac attagccgta ccgatggtgg ccgtatgctg atccacattg gtcgtaccaa
     1021 aaccctggtg agcaccgcgg gtgttgagaa ggcgctgagc ctgggcgtga ccaaactggt
     1081 tgaacgttgg atcagcgtga gcggtgttgc ggacgatccg aacaactacc tgttctgccg
     1141 tgttcgtaag aacggtgttg cggcgccgag cgcgaccagc cagctgagca cccgtgcgct
     1201 ggagggtatt tttgaagcga cccaccgtct gatctacggt gcgaaagacg atagcggcca
     1261 acgttatctg gcgtggagcg gtcacagcgc gcgtgtgggt gcggcgcgtg atatggcgcg
     1321 tgcgggcgtt agcattccgg aaatcatgca ggcgggtggc tggaccaacg tgaacattgt
     1381 tatgaactac attcgtaatc tggatagcga gaccggtgcg atggtgcgcc tgctggagga
     1441 tggtgattag taaggatcct ctagagtcga cctgcaggca tgcaagcttg gctgttttgg
     1501 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     1561 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     1621 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     1681 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1741 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatgt cgaatttgct ttcgaatttc
     1801 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     1861 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     1921 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     1981 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     2041 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     2101 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     2161 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     2221 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     2281 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
     2341 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     2401 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     2461 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     2521 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     2581 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     2641 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
     2701 gatttattta ttctgcgaag tgatcttccg tcacaggtat tttagctagc tcactcggtc
     2761 gctacgctcc gggcgtgaga ctgcggcggt agcggagtgt atactggctt actatgttgg
     2821 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     2881 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     2941 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     3001 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     3061 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     3121 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     3181 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     3241 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     3301 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     3361 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     3421 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     3481 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     3541 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     3601 agatcatctt attaatcaga taaaatattt ctagttttct actgaaccgc tctagatttc
     3661 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc c
//
