LOCUS       sequence-63931-e        4942 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 4942)
  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 4942)
  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: pSABAD332
FEATURES             Location/Qualifiers
     source          1..4942
                     /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             668..685
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(725..742)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      894..980
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1072..1099
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1117..1208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1209..1277,1278..2069)
                     /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(1427..1446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2111..2566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2198..2217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2408..2429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2677..3265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3166..3185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3419..3436
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3451..3591
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3517..3536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3677..3699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3838..3857
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4040..4918)
                     /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 catatgagaa taatatccaa
      361 aggagaactt gaattccatg aggtttcctc ggtaaggata atagattaca ataactgggt
      421 ctacgacctt gtaattccgg aaacccacaa cttcatagct cccaatggac ttgttctcca
      481 taatactcag ctaggtaccc agtacaaact tatcctgaac ggtaaaaccc tgaaaggtga
      541 aaccaccacc gaagctgttg acgctgctac cgcggaaaaa gttttcaaac agtacgctaa
      601 cgacaacggt gttgacggtg aatggaccta cgacgacgct accaaaacct acaccgtaac
      661 ggaattccat catcatcatc atcattaagc ttggctgttt tggcggatga gagaagattt
      721 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
      781 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
      841 gcgccgatgg tagtgtgggg tcccccatgc gagagtaggg aactgccagg catcaaataa
      901 aacgaaaggc tcagtgcaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
      961 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     1021 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     1081 tcctgacgga tggccttttt gcgtttctac aaactctttg tttatttttc taaatacatt
     1141 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1201 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1261 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1321 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1381 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1441 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1501 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1561 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1621 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1681 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1741 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1801 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1861 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1921 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1981 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2041 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2101 agattgattt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2161 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2221 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2281 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2341 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2401 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     2461 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2521 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt
     2581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2821 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3061 gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3301 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3361 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3421 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3481 gtgcggtatt tcacaccgga tatggtgcac tctcagtaca atctgctctg atgccgcata
     3541 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3601 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3661 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3721 cgcgcgaggc agcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     3781 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     3841 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     3901 gcgtccggcg tagaggatct aattctcatg tttgacagct tatcatcgat gcataatgtg
     3961 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa
     4021 ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac
     4081 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
     4141 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
     4201 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
     4261 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
     4321 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
     4381 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
     4441 acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg
     4501 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
     4561 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
     4621 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
     4681 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
     4741 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
     4801 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
     4861 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
     4921 ttttcatact cccgccattc ag
//
