LOCUS       sequence-62155-e        4824 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 4824)
  AUTHORS   Oeemig JS, Aranko AS, Djupsjobacka J, Heinamaki K, Iwai H
  TITLE     Solution structure of DnaE intein from Nostoc punctiforme: 
            structural basis for the design of a new split intein suitable for 
            site-specific chemical modification.
  JOURNAL   FEBS Lett. 2009 May 6;583(9):1451-6. doi: 
            10.1016/j.febslet.2009.03.058. Epub 2009 Apr 1.
  PUBMED    19344715
REFERENCE   2  (bases 1 to 4824)
  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"; doi: 
            "10.1016/j.febslet.2009.03.058"; journalName: "FEBS Lett"; date: 
            "2009-05-6- 6"; volume: "583"; issue: "9"; pages: "1451-6"
COMMENT     Sequence Label: pHYBAD94
FEATURES             Location/Qualifiers
     source          1..4824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             206..223
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(263..280)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      433..519
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      611..638
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        657..748
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(749..817,818..1609)
                     /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(967..986)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1651..2106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1738..1757)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1948..1969
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2217..2805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2706..2725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2959..2976
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2991..3131
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3057..3076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3217..3239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3378..3397
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3580..4458)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4485..4769
                     /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     4692..4711
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4796..4818
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatgggcttc atagcttcta attgtttcaa tggtacccag tacaaactta tcctgaacgg
       61 taaaaccctg aaaggtgaaa ccaccaccga agctgttgac gctgctaccg cggaaaaagt
      121 tttcaaacag tacgctaacg acaacggtgt tgacggtgaa tggacctacg acgacgctac
      181 caaaacctac accgtaacgg aattccatca tcatcatcat cattaagctt ggctgttttg
      241 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
      301 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
      361 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
      421 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      481 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
      541 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
      601 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
      661 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      721 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      781 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      841 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      901 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      961 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     1021 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1081 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1141 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1201 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1261 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     1321 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1381 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1441 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1501 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1561 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1621 agtttactca tatatacttt agattgattt acgcgccctg tagcggcgca ttaagcgcgg
     1681 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1741 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1801 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1861 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1921 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1981 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2041 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     2101 caatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2161 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2221 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2281 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2341 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2401 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2461 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2521 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2581 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     2641 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2701 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2761 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2821 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2881 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2941 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     3001 tattttctcc ttacgcatct gtgcggtatt tcacaccgga tatggtgcac tctcagtaca
     3061 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3121 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3181 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3241 tttcaccgtc atcaccgaaa cgcgcgaggc agcaaggaga tggcgcccaa cagtcccccg
     3301 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     3361 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     3421 ccggtgatgc cggccacgat gcgtccggcg tagaggatct aattctcatg tttgacagct
     3481 tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg
     3541 ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac
     3601 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
     3661 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
     3721 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
     3781 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
     3841 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
     3901 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
     3961 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
     4021 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
     4081 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
     4141 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
     4201 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
     4261 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
     4321 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     4381 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     4441 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     4501 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     4561 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
     4621 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
     4681 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
     4741 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagaa ataattttgt
     4801 ttaactttaa gaaggagata taca
//
