LOCUS       sequence-328229-        4265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains elements for building reporter plasmid for IBM. Built upon 
            pSB4A3. ECF20 reporter plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4265)
  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 4265)
  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: IBMc120
FEATURES             Location/Qualifiers
     source          1..4265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(331..425)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(455..1246,1247..1315))
                     /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     1078..1097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1316..1420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1485..1503
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1507..1550)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1553..1574
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             1646..1657
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2382..2453
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2469..2496
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2503..2523
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2524..2581
                     /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(2605..2622)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(2884..3834)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3882..4104
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt
       61 aaatggcact acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa
      121 aagcccgtca cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga
      181 ctttttgctg ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc
      241 ccgtgacagg tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg
      301 cttacccgtc ttactgtccc tagtgcttgg attctcacca ataaaaaacg cccggcggca
      361 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
      421 gagtccaagc gagctcgtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc atggtgccac
     1441 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     1501 ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc
     1561 ggccgcttct agaggcgcgg ataaaaattt catttgcccg cgacggattc cccgcccatc
     1621 tatcgttgaa cccatactag agattaaaga ggagaaatac tagatggtga gtaaaggaga
     1681 agctgtgatt aaagagttca tgcgcttcaa agttcacatg gagggttcta tgaacggtca
     1741 cgagttcgag atcgaaggcg aaggcgaggg ccgtccgtat gaaggcaccc agaccgccaa
     1801 actgaaagtg actaaaggcg gcccgctgcc tttttcctgg gacatcctga gcccgcaatt
     1861 tatgtacggt tctagggcgt tcatcaaaca cccagcggat atcccggact attataagca
     1921 gtcttttccg gaaggtttca agtgggaacg cgtaatgaat tttgaagatg gtggtgccgt
     1981 gaccgtcact caggacacct ccctggagga tggcaccctg atctataaag ttaaactgcg
     2041 tggtactaat tttccacctg atggcccggt gatgcagaaa aagacgatgg gttgggaggc
     2101 gtctaccgaa cgcttgtatc cggaagatgg tgtgctgaaa ggcgacatta aaatggccct
     2161 gcgcctgaaa gatggcggcc gctatctggc tgacttcaaa accacgtaca aagccaagaa
     2221 acctgtgcag atgcctggcg cgtacaatgt ggaccgcaaa ctggacatca cctctcataa
     2281 tgaagattat acggtggtag agcaatatga gcgctccgag ggtcgtcatt ctaccggtgg
     2341 catggatgaa ctatacaaat aataatacta gagccaggca tcaaataaaa cgaaaggctc
     2401 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     2461 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagta gcggccgctg
     2521 cagtccggca aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag
     2581 attacttcgc gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2641 gcggcgagcg gtatcagctc actcaaaggc ggtaatctcg aggttacatt gtcgatctgt
     2701 tcatggtgaa cagctttgaa tgcaccaaaa actcgtaaaa gctctgatgt atctatcttt
     2761 tttacaccgt tttcatctgt gcatatggac agttttccct ttgatatgta acggtgaaca
     2821 gttgttctac ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga
     2881 acctcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt
     2941 gagccatgag aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc
     3001 ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc
     3061 gttatgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat
     3121 ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat
     3181 caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt
     3241 taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt
     3301 attttcaagc attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg
     3361 agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt
     3421 ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata
     3481 aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt
     3541 tgtccactgg aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt
     3601 catcagctct ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat
     3661 catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc
     3721 aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa
     3781 gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct
     3841 caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt
     3901 acatgtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa
     3961 acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt
     4021 ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat
     4081 agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat
     4141 gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag
     4201 caccctcgca agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc
     4261 tgagt
//
