LOCUS       sequence-242897-        4916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Oth-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: pHBBAD168
FEATURES             Location/Qualifiers
     source          1..4916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..124
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     262..281
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(461..1339)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1366..1650
                     /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     1573..1592
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1677..1699
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2007..2024
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2064..2081)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2234..2320
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2412..2439
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2458..2549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2550..2618,2619..3410)
                     /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(2768..2787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3452..3907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3539..3558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3749..3770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4760..4777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(4792..4916,1..16)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4858..4877)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
       61 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
      121 gaggttttca ccgtcatcac cgaaacgcgc gaggcagaag gagatggcgc ccaacagtcc
      181 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
      241 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
      301 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc
      361 ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
      421 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
      481 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
      541 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
      601 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
      661 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
      721 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
      781 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
      841 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
      901 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
      961 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     1021 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     1081 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     1141 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     1201 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     1261 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     1321 cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc
     1381 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac
     1441 cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa
     1501 cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg
     1561 tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct
     1621 ttttatcgca actctctact gtttctccat acccgttttt ttgggctaga aataattttg
     1681 tttaacttta agaaggagat atacatatga gtgatattta ttgggatgaa atagtttcaa
     1741 aggaatatag tggtaaggaa gaagtgtttg acctcaccgt tcctggtttg cataattttg
     1801 ttgcaaataa tattattgtt cacaattcca ttggtaccca gtacaaactt atcctgaacg
     1861 gtaaaaccct gaaaggtgaa accaccaccg aagctgttga cgctgctacc gcggaaaaag
     1921 ttttcaaaca gtacgctaac gacaacggtg ttgacggtga atggacctac gacgacgcta
     1981 ccaaaaccta caccgtaacg gaattccatc atcatcatca tcattaagct tggctgtttt
     2041 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     2101 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     2161 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     2221 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2281 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     2341 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     2401 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt
     2461 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2521 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2581 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2641 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2701 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2761 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     2821 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2881 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2941 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3001 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3061 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     3121 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3181 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3241 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3301 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3361 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3421 aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg
     3481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3661 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3781 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3841 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3901 acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3961 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4681 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4741 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4801 gtattttctc cttacgcatc tgtgcggtat ttcacaccgg atatggtgca ctctcagtac
     4861 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtga
//
