LOCUS       sequence-63938-e        5704 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 5704)
  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 5704)
  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: pSABAD544
FEATURES             Location/Qualifiers
     source          1..5704
                     /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)"
     primer_bind     complement(377..405)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1411..1431
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1487..1504)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1656..1742
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1834..1861
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1879..1970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1971..2039,2040..2831)
                     /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(2189..2208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2873..3328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2960..2979)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3170..3191
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3439..4027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3928..3947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4181..4198
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4213..4353
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4279..4298)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4439..4461
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4600..4619
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4802..5680)
                     /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 catatggcta gcaaaggaga
      361 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
      421 caaattttct gtccgcggag agggtgaagg tgatgctaca aacggaaaac ttacccttaa
      481 atttatttgc actactggaa aactacctgt tccatggcca acactagtta ctactttatg
      541 tttaagctat gaaacggaaa tattgacagt agaatatgga ttattaccga ttggtaaaat
      601 tgtagaaaag cgcatcgaat gtactgttta tagcgttgat aataatggaa atatttatac
      661 acaacctgta gcacaatggc acgatcgcgg agaacaagag gtgtttgagt attgtttgga
      721 agatggttca ttgattcggg caacaaaaga ccataagttt atgactgttg atggtcaaat
      781 gttgccaatt gatgaaatat ttgaacgtga attggatttg atgcgggttg ataatttgcc
      841 gaatatcaaa atagccacac gtaaatattt aggcaaacaa aatgtctatg acattggagt
      901 tgagcgcgac cataattttg cactcaaaaa tggcttcata gccgcgggtt gttatggtgt
      961 tcaatgcttt tctagatatc cggatcatat gaaacggcat gactttttca agagtgccat
     1021 gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga actacaagac
     1081 gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaaaggtat
     1141 tgattttaaa gaagatggaa acattctcgg acacaaactc gagtacaact ataactcaca
     1201 caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg
     1261 ccacaacatt gaagatggat ccgttcaact agcagaccat tatcaacaaa atactccaat
     1321 tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat ctgccctttc
     1381 gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg
     1441 gttgacttaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     1501 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     1561 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     1621 ggtcccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtgca
     1681 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1741 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1801 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1861 ttgcgtttct acaaactctt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1921 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1981 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2041 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2101 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2161 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     2221 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2281 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2341 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2401 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2461 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2521 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2581 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2641 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2701 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2761 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2821 agcattggta actgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     2881 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2941 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3001 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3061 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3121 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3181 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     3241 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3301 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     3361 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3421 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3481 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3541 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3601 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3661 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3721 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3781 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     3841 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3901 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3961 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4021 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4081 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4141 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4201 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4261 gatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     4321 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     4381 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4441 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcaagga
     4501 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     4561 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     4621 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     4681 ctaattctca tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag
     4741 cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca
     4801 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     4861 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     4921 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     4981 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     5041 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
     5101 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     5161 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
     5221 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     5281 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     5341 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     5401 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     5461 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     5521 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     5581 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     5641 agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat
     5701 tcag
//
