LOCUS       sequence-9507-ex        4936 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 4936)
  AUTHORS   Muona M, Aranko AS, Iwai H
  TITLE     Segmental isotopic labelling of a multidomain protein by protein 
            ligation by protein trans-splicing.
  JOURNAL   Chembiochem. 2008 Dec 15. 9(18):2958-61.
  PUBMED    19031436
REFERENCE   2  (bases 1 to 4936)
  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: 
            "Chembiochem. 2008 Dec 15. 9(18):2958-61."
COMMENT     Sequence Label: pMMBAD16
FEATURES             Location/Qualifiers
     source          1..4936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(388..405)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      557..643
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      735..762
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        780..871
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(872..940,941..1732)
                     /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(1090..1109)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1774..2229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1861..1880)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2071..2092
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2340..2928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2829..2848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3082..3099
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3114..3254
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3180..3199)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3340..3362
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3501..3520
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3703..4581)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4608..4892
                     /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     4815..4834
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             join(4919..4936,1..5)
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ggagatatac atatggacca taattttgca ctcaaaaatg gcttcatagc ttctaattgt
       61 ttcaatggta ccctgggtgg gccggagcct gggccctatg cccaacccag cgtcaacact
      121 ccgctcccta acctccagaa tgggcccatt tatgccaggg ttatccagaa gcgagtccct
      181 aatgcctacg acaagacagc cttggctttg gaggtcggtg agctggtaaa ggttacgaag
      241 attaatgtga gtggtcagtg ggaaggggag tgtaatggca aacgaggtca cttcccattc
      301 acacatgtcc gtctgctgga tcaacagaat cccgatgagg acttcagcta agcttggctg
      361 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
      421 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
      481 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtccccca tgcgagagta
      541 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtgc aaagactggg cctttcgttt
      601 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
      661 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
      721 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
      781 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      841 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
      901 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
      961 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1021 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1081 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     1141 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1201 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1261 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1321 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1381 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     1441 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1501 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1561 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     1621 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     1681 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     1741 ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc gcattaagcg
     1801 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1861 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1921 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1981 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2041 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2101 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2161 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     2221 ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2461 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2701 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2941 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3001 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3061 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3121 cggtattttc tccttacgca tctgtgcggt atttcacacc ggatatggtg cactctcagt
     3181 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3241 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3301 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3361 ggttttcacc gtcatcaccg aaacgcgcga ggcagcaagg agatggcgcc caacagtccc
     3421 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     3481 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     3541 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctaattctc atgtttgaca
     3601 gcttatcatc gatgcataat gtgcctgtca aatggacgaa gcagggattc tgcaaaccct
     3661 atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca acttgacggc
     3721 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
     3781 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
     3841 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
     3901 gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg
     3961 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
     4021 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
     4081 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
     4141 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
     4201 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
     4261 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
     4321 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
     4381 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
     4441 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
     4501 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
     4561 atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag aaaccaattg
     4621 tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccaa
     4681 accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa
     4741 aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg
     4801 cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg
     4861 ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggcta gaaataattt
     4921 tgtttaactt taagaa
//
