LOCUS       sequence-65387-e        5686 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 5686)
  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 5686)
  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: pSABAD25-74
FEATURES             Location/Qualifiers
     source          1..5686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     111..130
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             215..237
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             257..274
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             284..301
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1370..1387)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1539..1625
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1717..1744
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1762..1853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1854..1922,1923..2714)
                     /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(2072..2091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2756..3211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2843..2862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3053..3074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3322..3910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3811..3830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4064..4081
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4096..4236
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4162..4181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4322..4344
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4483..4502
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4685..5563)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5590..5686,1..188)
                     /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)"
ORIGIN
        1 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
       61 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
      121 atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt
      181 ttctccatac ccgttttttt gggctagaaa taattttgtt taactttaag aaggagatat
      241 acccatgggc agcagccatc atcatcatca tcacagcagc ggcctggtgc cgcgcggcag
      301 cctaggacag tacaaactta tcctgaacgg taaaaccctg aaaggtgaaa ccaccaccga
      361 agctgttgac gctgctaccg cggaaaaagt tttcaaacag tacgctaacg acaacggtgt
      421 tgacggtgaa tggacctacg acgacgctac caaaacctac accgtaacgg aaggatccaa
      481 gtgcgtatca ggtgaaacac cagtttacct tgcggatggc aagacaataa aaataaagga
      541 tctatacagt tctgagagaa aaaaagaaga taacattgtt gaggctggtt cgggagaaga
      601 gataatacat ctaaaagatc ccattcaaat atattcttat gtggacggga ccatagtcag
      661 gagcagatca agacttctat acaagggcaa gagctcttat cttgtgagga tagaaactat
      721 tggcggaaga tcggtaagcg ttacaccagt tcacaaactc tttgtcctta cggaaaaggg
      781 tatcgaagag gttatggcct ccaacctaaa ggtaggcgac atgattgctg ctgtagcaga
      841 aagcgaatct gaagcaagag actgcggaat gagcgaggaa tgcgtgatgg aagcagaagt
      901 ttatacgtca cttgaagcga cattcgatag agtaaagtct atagcgtacg agaagggtga
      961 ttttgatgta tacgatcttt ccgtaccaga atacggcagg aactttatag gcggagaagg
     1021 acttctcgta cttcacaaca cggttatagg taccctgggt gggccggagc ctgggcccta
     1081 tgcccaaccc agcgtcaaca ctccgctccc taacctccag aatgggccca tttatgccag
     1141 ggttatccag aagcgagtcc ctaatgccta cgacaagaca gccttggctt tggaggtcgg
     1201 tgagctggta aaggttacga agattaatgt gagtggtcag tgggaagggg agtgtaatgg
     1261 caaacgaggt cacttcccat tcacacatgt ccgtctgctg gatcaacaga atcccgatga
     1321 ggacttcagc taagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     1381 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     1441 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     1501 tggggtcccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     1561 gcaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1621 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     1681 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     1741 tttttgcgtt tctacaaact ctttgtttat ttttctaaat acattcaaat atgtatccgc
     1801 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1861 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1921 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1981 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2041 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     2101 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2161 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2221 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2281 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2341 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     2401 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2461 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2521 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2581 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2641 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2701 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttacgcg
     2761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2941 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3061 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     3121 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3181 aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga tcctttttga
     3241 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3301 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3361 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3421 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3481 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3541 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3601 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3661 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     3721 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3781 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3841 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3901 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3961 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4021 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4081 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4141 ccggatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     4201 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     4261 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4321 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcaa
     4381 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     4441 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     4501 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     4561 gatctaattc tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg
     4621 aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta
     4681 ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact
     4741 cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa
     4801 aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg
     4861 cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga
     4921 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
     4981 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
     5041 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
     5101 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
     5161 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
     5221 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
     5281 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
     5341 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
     5401 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
     5461 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa
     5521 acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc
     5581 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
     5641 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaag
//
