LOCUS       sequence-372925-        4886 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 4886)
  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 4886)
  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: pSKBAD2
FEATURES             Location/Qualifiers
     source          1..4886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             58..80
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(415..432)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      585..671
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      763..790
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        809..900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(901..969,970..1761)
                     /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(1119..1138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1803..2258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1890..1909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2100..2121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2369..2957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2858..2877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3111..3128
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3143..3283
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3209..3228)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3369..3391
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3529..3548
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3728..4606)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(4633..4886,1..31)
                     /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     4840..4859
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag aaataatttt
       61 gtttaacttt aagaaggaga tatacatatg atcaaaatag ccacacgtaa atatttaggc
      121 aaacaaaatg tctatgacat tggagttgag cgcgaccata attttgcact caaaaatggc
      181 ttcatagctt ctaattgttt caatggtacc cagtacaaac ttatcctgaa cggtaaaacc
      241 ctgaaaggtg aaaccaccac cgaagctgtt gacgctgcta ccgcggaaaa agttttcaaa
      301 cagtacgcta acgacaacgg tgttgacggt gaatggacct acgacgacgc taccaaaacc
      361 tacaccgtaa cggaataagc ttggctgttt tggcggatga gagaagattt tcagcctgat
      421 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
      481 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
      541 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
      601 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
      661 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
      721 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
      781 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
      841 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      901 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      961 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1021 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1081 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1141 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1201 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1261 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1321 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1381 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1441 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1501 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1561 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1621 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1681 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1741 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1801 ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1861 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1921 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1981 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2041 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2101 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt
     2161 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2221 taacgcgaat tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc
     2281 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2341 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2401 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2461 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2521 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2581 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2641 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2701 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2761 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2821 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2881 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2941 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3001 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3061 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3121 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3181 tttcacaccg gatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3241 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3301 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3361 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3421 gcagaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     3481 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     3541 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     3601 gtagaggatc tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc tgtcaaatgg
     3661 acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg
     3721 ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     3781 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     3841 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
     3901 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     3961 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     4021 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     4081 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     4141 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     4201 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     4261 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     4321 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     4381 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     4441 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     4501 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     4561 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     4621 cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
     4681 tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac
     4741 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
     4801 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
     4861 taagattagc ggatcctacc tgacgc
//
