LOCUS       sequence-242763-        4916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Charge-Introduced NpuDnaBC39 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4916)
  AUTHORS   Beyer HM, Mikula KM, Li M, Wlodawer A, Iwai H
  TITLE     The crystal structure of the naturally split gp41-1 intein guides 
            the engineering of orthogonal split inteins from cis-splicing 
            inteins.
  JOURNAL   FEBS J. 2019 Oct 30. doi: 10.1111/febs.15113.
  PUBMED    31665813
REFERENCE   2  (bases 1 to 4916)
  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: "FEBS J. 
            2019 Oct 30. doi: 10.1111/febs.15113."
COMMENT     Sequence Label: pHBBAD113
FEATURES             Location/Qualifiers
     source          1..4916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     215..232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    247..387
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(313..332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     473..495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     633..652
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(832..1710)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1737..2021
                     /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     1944..1963
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2048..2070
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2378..2395
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2435..2452)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2605..2691
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2783..2810
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2829..2920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2921..2989,2990..3781)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3139..3158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3823..4278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3910..3929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4120..4141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      join(4389..4916,1..61)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4878..4897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
       61 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      121 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      181 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      241 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accggatatg
      301 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
      361 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
      421 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
      481 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg
      541 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
      601 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
      661 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca
      721 tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag cagggattct
      781 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
      841 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
      901 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
      961 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     1021 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     1081 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     1141 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     1201 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     1261 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     1321 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     1381 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     1441 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     1501 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     1561 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     1621 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     1681 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga
     1741 aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc
     1801 gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc
     1861 catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta
     1921 tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc
     1981 tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag
     2041 aaataatttt gtttaacttt aagaaggaga tatacatatg agtgatattt attgggatga
     2101 aatagttaaa aaggaatata gtggtgagga agaagtgttt gacctcaccg ttcctggttt
     2161 gcataatttt gttgcaaata atattattgt tcacaattcc attggtaccc agtacaaact
     2221 tatcctgaac ggtaaaaccc tgaaaggtga aaccaccacc gaagctgttg acgctgctac
     2281 cgcggaaaaa gttttcaaac agtacgctaa cgacaacggt gttgacggtg aatggaccta
     2341 cgacgacgct accaaaacct acaccgtaac ggaattccat catcatcatc atcattaagc
     2401 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     2461 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     2521 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     2581 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2641 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     2701 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     2761 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     2821 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2881 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2941 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3001 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3061 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3121 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     3181 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3241 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3301 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3361 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3421 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     3481 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3541 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3601 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3661 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3721 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3781 actgtcagac caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg
     3841 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3901 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3961 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4021 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4081 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4141 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4201 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4261 tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4321 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4381 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4441 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4501 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4561 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4621 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4681 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4741 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4801 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4861 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcg
//
