LOCUS       sequence-124229-        4194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cisRAJ31-cat, a cis-repressed mRNA regulated by RAJ31..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4194)
  AUTHORS   Rostain W, Shen S, Cordero T, Rodrigo G, Jaramillo A
  TITLE     Engineering a Circular Riboregulator in Escherichia coli.
  JOURNAL   Biodes Res. 2020 Sep 12;2020:1916789. doi: 10.34133/2020/1916789. 
            eCollection 2020.
  PUBMED    37849901
REFERENCE   2  (bases 1 to 4194)
  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: "Biodes 
            Res."; date: "2020-09-12"; volume: "2020:1916789. doi"; pages: " 
            10.34133/2020/1916789. eCollection 2020"
COMMENT     Sequence Label: cisRAJ31-cat
FEATURES             Location/Qualifiers
     source          1..4194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..816)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     724..743
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     808..827
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1174..2124)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2172..2394
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2886..2980)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(3136..3165)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    3212..3231
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     protein_bind    3265..3281
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             3328..3339
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3346..4005
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3412..3431)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_feature    4009..4029
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4065..4096
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
       61 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      121 aagaatggca aggtcctggt aacggtctgc gattccgacc cgtccaacat caatacaacc
      181 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      241 tgaatccggt gagaatggca agagcttgtg catttctttc cagacttgtt caacaggcca
      301 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca tgcgtgattg
      361 cgcctgagca agacgaaata cacgatcgct gttaaaagga caattacaaa caggaatcga
      421 atgtaaccgg cgcaggaaca cggccagcgc atcaacaata ttttcacctg aatcaggata
      481 ttcttctaat acctggaagg ctgttttccc aggaatcgcg gtggtgagta accacgcatc
      541 atcaggagta cggataaaat gcttgatggt cgggagaggc ataaactccg tcagccagtt
      601 gagacggacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
      661 caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac
      721 attatcgcga gcccatttat acccatataa atcagcgtcc atgttggagt ttaagcgcgg
      781 acgggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
      841 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
      901 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
      961 tcagctctag tagttacatt gtcgatctgt tcatggtgaa cagctttgaa tgcaccaaaa
     1021 actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac
     1081 agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat
     1141 gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat
     1201 gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata
     1261 cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt
     1321 taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt
     1381 tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc
     1441 catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc
     1501 aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca
     1561 ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc
     1621 atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa
     1681 ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata
     1741 ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc
     1801 taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatcccaa agcctttaac
     1861 caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac
     1921 accataagca ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga
     1981 taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca
     2041 taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc
     2101 tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata
     2161 ccaattgaga tgggctagtc aatgataatt acatgtcctt ttcctttgag ttgtgggtat
     2221 ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa
     2281 ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat
     2341 aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact
     2401 ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa
     2461 cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa
     2521 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
     2581 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
     2641 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
     2701 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
     2761 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
     2821 aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctagtg
     2881 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     2941 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgtaaacttg
     3001 gtctgacagc tctagctccg gcaaaaaaac gggcaaggtg tcaccaccct gccctttttc
     3061 tttaaaaccg aaaagattac ttcgcgtttg ccacctgacg tctaagaaaa ggaatattca
     3121 gcaatttgcc cgtgccgaag aaaggcccac ccgtgaaggt gagccagtga gttgattgct
     3181 acgtaattag ttagttagcc cttagtgact cgaattcgcg gccgcttcta ggatgcatat
     3241 aaatgtgagc ggataacatt gacattgtga gcggataaca agatactgag cacagtggat
     3301 atgctttacc tcttggtgag acgacataaa gaggagaaac ataatatgga gaaaaaaatc
     3361 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     3421 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     3481 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     3541 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     3601 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     3661 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     3721 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     3781 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     3841 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     3901 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     3961 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaataata ctagtagcgg
     4021 ccgctgcagg agtcactaag ggttagttag ttagattagc agaaagtcaa aagcctccga
     4081 ccggaggctt ttgactaaaa cttcccttgg ggttatcatt ggggctcact caaaggcggt
     4141 aatcagataa aaaaaatcct tagctttcgc taaggatgat ttctgctagt atta
//
