LOCUS       sequence-314129-        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(318..412)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(442..1233,1234..1302))
                     /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     1065..1084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1303..1407)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1472..1490
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1494..1537)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1540..1561
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             1633..1644
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2369..2440
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2456..2483
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2490..2510
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2511..2568
                     /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(2592..2609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(2871..3821)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3869..4091
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca
       61 ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg
      121 gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc
      181 agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca
      241 ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta
      301 ctgtccctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
      361 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
      421 ctcgtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      481 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
      541 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
      601 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      661 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      721 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      781 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      841 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      901 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      961 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1021 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1081 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1141 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1201 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1261 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1381 gaaaaataaa caaatagggg ttccgcgcac atttcccatg gtgccacctg acgtctaaga
     1441 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag
     1501 ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgcggc cgcttctaga
     1561 ggcgcggata aaaatttcat ttgcccgcga cggattcccc gcccatctat cgttgaaccc
     1621 atactagaga ttaaagagga gaaatactag atggtgagta aaggagaagc tgtgattaaa
     1681 gagttcatgc gcttcaaagt tcacatggag ggttctatga acggtcacga gttcgagatc
     1741 gaaggcgaag gcgagggccg tccgtatgaa ggcacccaga ccgccaaact gaaagtgact
     1801 aaaggcggcc cgctgccttt ttcctgggac atcctgagcc cgcaatttat gtacggttct
     1861 agggcgttca tcaaacaccc agcggatatc ccggactatt ataagcagtc ttttccggaa
     1921 ggtttcaagt gggaacgcgt aatgaatttt gaagatggtg gtgccgtgac cgtcactcag
     1981 gacacctccc tggaggatgg caccctgatc tataaagtta aactgcgtgg tactaatttt
     2041 ccacctgatg gcccggtgat gcagaaaaag acgatgggtt gggaggcgtc taccgaacgc
     2101 ttgtatccgg aagatggtgt gctgaaaggc gacattaaaa tggccctgcg cctgaaagat
     2161 ggcggccgct atctggctga cttcaaaacc acgtacaaag ccaagaaacc tgtgcagatg
     2221 cctggcgcgt acaatgtgga ccgcaaactg gacatcacct ctcataatga agattatacg
     2281 gtggtagagc aatatgagcg ctccgagggt cgtcattcta ccggtggcat ggatgaacta
     2341 tacaaataat aatactagag ccaggcatca aataaaacga aaggctcagt cgaaagactg
     2401 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     2461 accttcgggt gggcctttct gcgtttatat actagtagcg gccgctgcag tccggcaaaa
     2521 aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt
     2581 atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2641 tcagctcact caaaggcggt aatctcgagg ttacattgtc gatctgttca tggtgaacag
     2701 ctttaaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
     2761 catctgtgca tatggacagt tttccctttg atatctaacg gtgaacagtt gttctacttt
     2821 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     2881 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     2941 gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     3001 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt acgtaggtag
     3061 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     3121 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     3181 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     3241 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     3301 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     3361 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     3421 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     3481 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     3541 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     3601 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     3661 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     3721 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     3781 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     3841 tgattttaat cactatacca attgagatgg gctagtcaat gataattaca tgtccttttc
     3901 ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     3961 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     4021 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     4081 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     4141 tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc
     4201 tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct
     4261 ttttc
//
