LOCUS       sequence-63865-e        5110 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 5110)
  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 5110)
  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: pSABAD219
FEATURES             Location/Qualifiers
     source          1..5110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             79..101
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             601..618
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(658..675)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      827..913
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1005..1032
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1050..1141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1142..1210,1211..2002)
                     /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(1360..1379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2044..2499
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2131..2150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2341..2362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2610..3198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3099..3118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3352..3369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3384..3524
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3450..3469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3610..3632
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3771..3790
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3973..4851)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(4878..5110,1..52)
                     /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     5085..5104
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt
       61 ttttgggcta gaaataattt tgtttaactt taagaaggag atatacatat gaatggaaat
      121 atttatacac aacctgtagc acaatggcac gatcgcggag aacaagaggt gtttgagtat
      181 tgtttggaag atggttcatt gattcgggca acaaaagacc ataagtttat gactgttgat
      241 ggtcaaatgt tgccaattga tgaaatattt gaacgtgaat tggatttgat gcgggttgat
      301 aatttgccga atatcaaaat agccacacgt aaatatttag gcaaacaaaa tgtctatgac
      361 attggagttg agcgcgacca taattttgca ctcaaaaatg gcttcatagc ttctaattgt
      421 ttcaatggta cccagtacaa acttatcctg aacggtaaaa ccctgaaagg tgaaaccacc
      481 accgaagctg ttgacgctgc taccgcggaa aaagttttca aacagtacgc taacgacaac
      541 ggtgttgacg gtgaatggac ctacgacgac gctaccaaaa cctacaccgt aacggaattc
      601 catcatcatc atcatcatta agcttggctg ttttggcgga tgagagaaga ttttcagcct
      661 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
      721 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
      781 tggtagtgtg gggtccccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
      841 ggctcagtgc aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      901 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
      961 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     1021 ggatggcctt tttgcgtttc tacaaactct ttgtttattt ttctaaatac attcaaatat
     1081 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1141 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1201 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1261 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1321 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1381 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1441 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1501 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1561 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1621 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1681 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1741 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1801 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1861 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1921 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1981 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2041 tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2101 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2161 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2221 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2281 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2341 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     2401 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2461 ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc
     2521 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2581 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2641 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2701 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2761 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2821 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2881 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2941 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3001 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3061 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3121 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3181 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3241 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3301 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3361 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3421 atttcacacc ggatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3481 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3541 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3601 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3661 ggcagcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     3721 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     3781 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     3841 gcgtagagga tctaattctc atgtttgaca gcttatcatc gatgcataat gtgcctgtca
     3901 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     3961 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     4021 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     4081 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     4141 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     4201 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     4261 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     4321 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     4381 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     4441 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     4501 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     4561 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     4621 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     4681 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     4741 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     4801 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     4861 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     4921 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     4981 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     5041 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     5101 atccataaga
//
