LOCUS       sequence-63929-e        4948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4948)
  AUTHORS   Aranko AS, Oeemig JS, Kajander T, Iwai H
  TITLE     Intermolecular domain swapping induces intein-mediated protein 
            alternative splicing.
  JOURNAL   Nat Chem Biol. 2013 Aug 25. doi: 10.1038/nchembio.1320.
  PUBMED    23974115
REFERENCE   2  (bases 1 to 4948)
  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 Chem 
            Biol. 2013 Aug 25. doi: 10.1038/nchembio.1320."
COMMENT     Sequence Label: pSABAD331
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..287
                     /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     210..229
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             314..336
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             674..691
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(731..748)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      900..986
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1078..1105
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1123..1214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1215..1283,1284..2075)
                     /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(1433..1452)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2117..2572
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2204..2223)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2414..2435
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2683..3271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3172..3191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3425..3442
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3457..3597
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3523..3542)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3683..3705
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3844..3863
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4046..4924)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
       61 cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
      121 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
      181 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
      241 ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgtttttttg
      301 ggctagaaat aattttgttt aactttaaga aggagatata catatggaag tttatacgtc
      361 acttgaagcg acattcgata gagtaaagtc tatagcgtac gagaagggtg attttgatgt
      421 atacgatctt tccgtaccag aatacggcag gaactttata ggcggagaag gacttctcgt
      481 acttcacaac acggttatag gtacccagta caaacttatc ctgaacggta aaaccctgaa
      541 aggtgaaacc accaccgaag ctgttgacgc tgctaccgcg gaaaaagttt tcaaacagta
      601 cgctaacgac aacggtgttg acggtgaatg gacctacgac gacgctacca aaacctacac
      661 cgtaacggaa ttccatcatc atcatcatca ttaagcttgg ctgttttggc ggatgagaga
      721 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
      781 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
      841 gccgtagcgc cgatggtagt gtggggtccc ccatgcgaga gtagggaact gccaggcatc
      901 aaataaaacg aaaggctcag tgcaaagact gggcctttcg ttttatctgt tgtttgtcgg
      961 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     1021 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     1081 aggccatcct gacggatggc ctttttgcgt ttctacaaac tctttgttta tttttctaaa
     1141 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1201 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1261 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1321 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1381 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1441 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1501 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1561 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1621 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1681 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1741 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     1801 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1861 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1921 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1981 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2041 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2101 tactttagat tgatttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2161 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2221 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2281 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2341 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2401 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     2461 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2521 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttaaaaggat
     2581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2701 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2761 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2821 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2881 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2941 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3001 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3061 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3121 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3181 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3241 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3301 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3361 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3421 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     3481 gcatctgtgc ggtatttcac accggatatg gtgcactctc agtacaatct gctctgatgc
     3541 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3601 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3661 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3721 ccgaaacgcg cgaggcagca aggagatggc gcccaacagt cccccggcca cggggcctgc
     3781 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     3841 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     3901 cacgatgcgt ccggcgtaga ggatctaatt ctcatgtttg acagcttatc atcgatgcat
     3961 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     4021 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     4081 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     4141 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     4201 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     4261 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     4321 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     4381 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     4441 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     4501 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     4561 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     4621 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     4681 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     4741 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     4801 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     4861 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     4921 ccatactttt catactcccg ccattcag
//
