LOCUS       sequence-8261-ex        4888 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 4888)
  AUTHORS   Iwai H, Zuger S, Jin J, Tam PH
  TITLE     Highly efficient protein trans-splicing by a naturally split DnaE 
            intein from Nostoc punctiforme.
  JOURNAL   FEBS Lett. 2006 Mar 20. 580(7):1853-8.
  PUBMED    16516207
REFERENCE   2  (bases 1 to 4888)
  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: "FEBS Lett. 
            2006 Mar 20. 580(7):1853-8."
COMMENT     Sequence Label: pJJBAD78
FEATURES             Location/Qualifiers
     source          1..4888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1312..1334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1669..1686)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1838..1924
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2016..2043
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2061..2152
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2153..2221,2222..3013)
                     /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(2371..2390)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3055..3510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3142..3161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3352..3373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3621..4209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4110..4129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4363..4380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4395..4535
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4461..4480)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4621..4643
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4782..4801
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagaaataa ttttgtttaa
     1321 ctttaagaag gagatataca tatggttaaa gttatcggtc gtcgatccct gggtgtgcaa
     1381 cgcatctttg atatcggtct gccgcaagac cataattttc tgctggctaa tggtgctatc
     1441 gccgccaact gtatgaacgg tacccagtac aaacttatcc tgaacggtaa aaccctgaaa
     1501 ggtgaaacca ccaccgaagc tgttgacgct gctaccgcgg aaaaagtttt caaacagtac
     1561 gctaacgaca acggtgttga cggtgaatgg acctacgacg acgctaccaa aacctacacc
     1621 gtaacggaat aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     1681 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     1741 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     1801 ggggtccccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtg
     1861 caaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     1921 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     1981 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     2041 ttttgcgttt ctacaaactc tttgtttatt tttctaaata cattcaaata tgtatccgct
     2101 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2161 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2221 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2281 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2341 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     2401 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2461 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2521 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2581 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2641 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     2701 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2761 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2821 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2881 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2941 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3001 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc
     3061 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3121 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3181 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3241 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3301 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3361 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     3421 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3481 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat
     3541 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3601 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3661 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3721 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3781 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3841 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3901 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3961 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     4021 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4081 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4141 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4201 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4261 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4321 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4381 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4441 cggatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4501 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4561 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4621 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaag
     4681 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     4741 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     4801 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     4861 atctaattct catgtttgac agcttatc
//
