LOCUS       sequence-326341-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Type IIS restriction site free elements for expressing 
            protein to be split in IBM. TetR expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  AUTHORS   Ho TYH, Shao A, Lu Z, Savilahti H, Menolascina F, Wang L, Dalchau N,
            Wang B
  TITLE     A systematic approach to inserting split inteins for Boolean logic 
            gate engineering and basal activity reduction.
  JOURNAL   Nat Commun. 2021 Apr 13;12(1):2200. doi: 10.1038/s41467-021-22404-9.
  PUBMED    33850130
REFERENCE   2  (bases 1 to 4743)
  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: "Nat 
            Commun."; date: "2021-04-13"; volume: "12(1):2200. doi"; pages: " 
            10.1038/s41467-021-22404-9"
COMMENT     Sequence Label: IBMc226
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(3..71,72..863)
                     /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     complement(221..240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(990..1019)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    1066..1087
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1088..1966)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1993..2277
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2200..2219
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2314..2937
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2387..2406)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    2999..3019
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3055..3086
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      3597..4141
                     /direction=RIGHT
                     /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     4042..4061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4258..4275
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
       61 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      121 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      181 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      241 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      301 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      361 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      421 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      481 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      541 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      601 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      661 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat
      721 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
      781 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
      841 cctcactgat taagcattgg taatactagc tccggcaaaa aaacgggcaa ggtgtcacca
      901 ccctgccctt tttctttaaa accgaaaaga ttacttcgcg tttgccacct gacgtctaag
      961 aaaaggaata ttcagcaatt tgcccgtgcc gaagaaaggc ccacccgtga aggtgagcca
     1021 gtgagttgat tgctacgtaa ttagttagtt agcccttagt gactcgaatt cgcggccgct
     1081 tctagagtta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     1141 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     1201 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
     1261 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     1321 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     1381 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     1441 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     1501 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     1561 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     1621 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     1681 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     1741 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     1801 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     1861 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     1921 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     1981 cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
     2041 tttactggca cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac
     2101 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
     2161 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
     2221 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccattac
     2281 tagagggaag agagtcaatt cagggtggtg aatatgtcca gattagataa aagtaaagtg
     2341 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa
     2401 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg
     2461 gctttgctcg acgccttagc cattgagatg ttagataggc accatactca cttttgccct
     2521 ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct
     2581 ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag
     2641 tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat
     2701 gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa
     2761 gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta
     2821 ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc
     2881 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcctgatac
     2941 tagagaacgc atgagaaagc ccccggaaga tcaccttccg ggggcttttt tattgcgcta
     3001 ctagtagcgg ccgctgcagg agtcactaag ggttagttag ttagattagc agaaagtcaa
     3061 aagcctccga ccggaggctt ttgactaaaa cttcccttgg ggttatcatt ggggctcact
     3121 caaaggcggt aatcagataa aaaaaatcct tagctttcgc taaggatgat ttctgctaga
     3181 gatggaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggatc tcgcggtatc
     3301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3541 ccttaacgtg agttttcgtt ccactgagcg tcagacccct taataagatg atcttcttga
     3601 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     3661 ggttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag
     3721 cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta
     3781 actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg
     3841 ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg
     3901 tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg
     3961 agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga
     4021 gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4081 ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa
     4141 aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg
     4201 aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc
     4261 gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt
     4321 gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat
     4381 agtaagccag tatacactcc gctagcgctg aggtctgcct cgtgaagaag gtgttgctga
     4441 ctcataccag gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat
     4501 gagagctttg ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg
     4561 gtctgcgttg tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat
     4621 tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     4681 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgtttacta
     4741 gag
//
