LOCUS       sequence-7154-ex        4879 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 4879)
  AUTHORS   Zuger S, Iwai H
  TITLE     Intein-based biosynthetic incorporation of unlabeled protein tags 
            into isotopically labeled proteins for NMR studies.
  JOURNAL   Nat Biotechnol. 2005 Jun . 23(6):736-40.
  PUBMED    15908942
REFERENCE   2  (bases 1 to 4879)
  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 
            Biotechnol. 2005 Jun . 23(6):736-40."
COMMENT     Sequence Label: pSFBAD09
FEATURES             Location/Qualifiers
     source          1..4879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(186..1064)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1091..1375
                     /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     1298..1317
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1402..1424
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1555..1710
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(1750..1767)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1919..2005
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2097..2124
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2142..2233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2234..2302,2303..3094)
                     /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(2452..2471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3136..3591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3223..3242)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3433..3454
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3702..4290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4191..4210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4444..4461
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4476..4616
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4542..4561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4702..4724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(4863..4879,1..3)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
       61 ggatctaatt ctcatgtttg acagcttatc atcgatgcat aatgtgcctg tcaaatggac
      121 gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt
      181 accaattatg acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac
      241 tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca
      301 aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc
      361 gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg
      421 aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca
      481 aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc
      541 atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc
      601 agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc
      661 ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg
      721 gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc
      781 cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc
      841 gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc
      901 ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata
      961 aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta
     1021 aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg
     1081 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
     1141 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
     1201 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
     1261 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
     1321 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg
     1381 tttttttggg ctagaaataa ttttgtttaa ctttaagaag gagatataca tatggttaaa
     1441 gttatcggtc gtcgatccct gggtgtgcaa cgcatctttg atatcggtct gccgcaagac
     1501 cataattttc tgctggctaa tggtgctatc gccgccaact gttttaacaa aggtaccaca
     1561 aatcctggtg tatccgcttg gcaggtcaac acagcttata ctgcgggaca attggtcaca
     1621 tataacggca agacgtataa atgtttgcag ccccacacct ccttggcagg atgggaacca
     1681 tccaacgttc ctgccttgtg gcagcttcaa taagcttggc tgttttggcg gatgagagaa
     1741 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     1801 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     1861 ccgtagcgcc gatggtagtg tggggtcccc catgcgagag tagggaactg ccaggcatca
     1921 aataaaacga aaggctcagt gcaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     1981 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     2041 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     2101 ggccatcctg acggatggcc tttttgcgtt tctacaaact ctttgtttat ttttctaaat
     2161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2221 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2281 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2341 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2401 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2461 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2521 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2581 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2641 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2701 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2761 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2821 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2881 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2941 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3001 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3061 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3121 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3181 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3241 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3301 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3361 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3421 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     3481 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3541 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc
     3601 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3841 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4081 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4321 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4381 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4441 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4501 catctgtgcg gtatttcaca ccggatatgg tgcactctca gtacaatctg ctctgatgcc
     4561 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     4621 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4681 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4741 cgaaacgcgc gaggcagcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     4801 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     4861 tcggtgatgt cggcgatat
//
