LOCUS       sequence-329041-        4689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Carries Golden Gate substitution insert for IBM with gp41-1 intein. 
            PPhlF as C-lobe promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4689)
  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 4689)
  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: IBMc460
FEATURES             Location/Qualifiers
     source          1..4689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(51..842,843..911))
                     /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     674..693
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(912..1016)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1084..1102
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1106..1149)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1152..1173
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1647..2306)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2221..2240
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3346..3365)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_RNA        3559..3608
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_feature    3784..3804
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(3810..3827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3981..4569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4061..4080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
       61 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      121 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      181 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      241 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      301 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      361 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      421 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      481 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      541 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      601 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      661 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      721 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      781 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      841 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      901 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      961 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1021 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1081 aaaaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag
     1141 gatgatttct ggaattcgcg gccgcttcta gaggaagacg ctggatattg tctggatctg
     1201 aaaacccagg ttcagacacc gcagggtatg aaagaaattt caaatattca ggtgggtgat
     1261 ctggttctga gcaataccgg ttataatgaa gtgctgaatg tgtttccgaa aagcaaaaaa
     1321 aaaagctata aaatcaccct ggaagatggc aaagaaatca tttgtagcga agaacacctg
     1381 tttccgaccc agaccggtga aatgaatatt agcggtggtc tgaaagaagg tatgtgcctg
     1441 tatgttaaag aataatacta gagttgagaa gagaaaagaa aaccgccgat cctgtccacc
     1501 gcattactgc aaggtagtgg acaagaccgg cggtcttaag ttttttggct gaatactaga
     1561 gcctttggtc gaaaaaaaaa gcccgcactg tcaggtgcgg gcttttttct gtgtttccta
     1621 ctagagcaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
     1681 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1741 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1801 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1861 ggattggctg aaacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1921 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1981 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     2041 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     2101 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     2161 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     2221 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     2281 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     2341 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     2401 tgccctcggt accaaattcc agaaaagagg cctcccgaaa ggggggcctt ttttcgtttt
     2461 ggtccaatgg cggcgcgcca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
     2521 cccggaagag agtcaattca tgggggtgaa tatggcacgt accccgagcc gtagcagcat
     2581 tggtagcctg cgtagtccgc atacccataa agcaattctg accagcacca ttgaaatcct
     2641 gaaagaatgt ggttatagcg gtctgagcat tgaaagcgtg gcacgtcgcg ccggtgcagg
     2701 caaaccgacc atttatcgtt ggtggaccaa caaagcagca ctgattgccg aagtgtatga
     2761 aaatgaaatc gaacaggtac gtaaatttcc ggatttgggt agctttaaag ccgatctgga
     2821 ttttctgctg cataatctgt ggaaagtttg gcgtgaaacc atttgtggtg aagcatttcg
     2881 ttgtgttatt gcagaagcac agttggaccc tgtaaccctg acccaactga aagatcagtt
     2941 tatggaacgt cgtcgtgaga taccgaaaaa actggttgaa gatgccatta gcaatggtga
     3001 actgccgaaa gatatcaatc gtgaactgct gctggatatg atttttggtt tttgttggta
     3061 tcgcctgctg accgaacagt tgaccgttga acaggatatt gaagaattta ccttcctgct
     3121 gattaatggt gtttgtccgg gtacacagtg ttgataagga tcctaattgg taacgaatca
     3181 gacaattgac ggctcgaggg agtagcatag ggtttgcaga atccctgctt cgtccatttg
     3241 acaggcacat tatgcatcga tgataagctg tcaaacatga gcagatcctc tacgccggac
     3301 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     3361 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcaaatat tttatctgcc
     3421 aattattgaa ggcctcccta acggggggcc tttttttgtt tctggtctcc cgcttaacga
     3481 tcgttggctg cgacgtacgg tggaatctga ttcgttacca attgacatga tacgaaacgt
     3541 accgtatcgt taaggtagct gtcaccggat gtgctttccg gtctgatgag tccgtgagga
     3601 cgaaacagcc tctacaaata attttgttta atactagagt cacacaggaa agtactagat
     3661 gatgctgaaa aaaatcctga aaatcgagga actggatgaa cgcgaactga ttgatattga
     3721 agttagcggt aaccacctgt tctatgccaa tgatattctg acccataatt cgtcaagaag
     3781 agctactagt agcggccgct gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3961 aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat
     4021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4381 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaaagaaaac
     4621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4681 aattaaaaa
//
