LOCUS       sequence-100918-        4826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TvoVMAC7 intein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4826)
  AUTHORS   Aranko AS, Oeemig JS, Zhou D, Kajander T, Wlodawer A, Iwai H
  TITLE     Structure-based engineering and comparison of novel split inteins 
            for protein ligation.
  JOURNAL   Mol Biosyst. 2014 May;10(5):1023-34. doi: 10.1039/c4mb00021h.
  PUBMED    24574026
REFERENCE   2  (bases 1 to 4826)
  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"; doi: "10"; journalName: 
            "Mol Biosyst"; date: "2014-05"; volume: "10"; issue: "5"; pages: 
            "1023-34"
COMMENT     Sequence Label: pHYBAD177
FEATURES             Location/Qualifiers
     source          1..4826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             27..49
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             264..281
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(321..338)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      491..577
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      669..696
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        714..805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(806..874,875..1666)
                     /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(1024..1043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1708..2163
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1795..1814)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2005..2026
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2274..2862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2763..2782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3016..3033
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3048..3188
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3114..3133)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3274..3296
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3435..3454
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3637..4515)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4542..4826
                     /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     4749..4768
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 acccgttttt ttgggctaga aataattttg tttaacttta agaaggagat atacatatgg
       61 gacttctcgt acttcacaac acggttatag gtacccagta caaacttatc ctgaacggta
      121 aaaccctgaa aggtgaaacc accaccgaag ctgttgacgc tgctaccgcg gaaaaagttt
      181 tcaaacagta cgctaacgac aacggtgttg acggtgaatg gacctacgac gacgctacca
      241 aaacctacac cgtaacggaa ttccatcatc atcatcatca ttaagcttgg ctgttttggc
      301 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
      361 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
      421 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
      481 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
      541 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
      601 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
      661 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctctttgttt
      721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1021 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1321 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     1381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1681 ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta agcgcggcgg
     1741 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1801 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1861 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1921 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1981 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2041 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2101 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2161 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2221 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2281 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2341 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2401 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2461 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2521 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2581 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2641 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     2701 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2761 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2821 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2881 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2941 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3001 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3061 tttctcctta cgcatctgtg cggtatttca caccggatat ggtgcactct cagtacaatc
     3121 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3181 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3241 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3301 caccgtcatc accgaaacgc gcgaggcagc aaggagatgg cgcccaacag tcccccggcc
     3361 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     3421 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     3481 gtgatgccgg ccacgatgcg tccggcgtag aggatctaat tctcatgttt gacagcttat
     3541 catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta
     3601 ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga cggctacatc
     3661 attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt
     3721 aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat
     3781 aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca
     3841 gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa
     3901 gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat
     3961 gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc
     4021 ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg
     4081 cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg
     4141 cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc
     4201 atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg
     4261 atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca
     4321 aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat
     4381 ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat
     4441 cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt
     4501 ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata
     4561 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt
     4621 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg
     4681 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac
     4741 actttgctat gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt
     4801 atcgcaactc tctactgttt ctccat
//
