LOCUS       sequence-247669-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DpnII restriction endonuclease in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  TITLE     Bacterial expression plasmid for DpnII restriction endonuclease
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6245)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBAD33.1-DpnII
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..299
                     /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     222..241
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             359..381
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1293..1310)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1463..1549
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1641..1668
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1687..1778
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2236..2691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2323..2342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2533..2554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3247..3906)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3821..3840
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3907..4009)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4401..4418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4535..5080)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4615..4634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5156..5175
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5355..6233)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
       61 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
      121 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
      181 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
      241 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
      301 cccgtttttt tgggctagcg aattcgagct cggtacccgg ggatcctcta gaaataattt
      361 tgtttaactt taagaaggag atatacatat gaagcaaacg cgcaattttg atgaatggct
      421 gagcacgatg accgacacgg tcgccgactg gacctactat actgattttc caaaagttta
      481 caagaacgtg tctagcatca aagtcgctct taacattatg aacagtttga ttggtagtaa
      541 gaacattcaa gaagattttc ttgacctgta ccaaaactac ccggaaattc tgaaagtagt
      601 cccccttttg atcgcaaagc gtttgcgtga tacgattatc gtaaaagacc ctatcaagga
      661 cttttacttc gatttttcaa aacgtaacta tagtattgag gagtacacga tgttccttga
      721 aaagtcgggg atctttgatc tgctgcaaaa tcacttagtt agcaatttag ttgattacgt
      781 gactggggtc gaggtcggta tggacaccaa cgggcgtaag aaccgcacag gagacgcgat
      841 ggaaaacatt gtgcaatcgt acttggaagc cgaaggttat atccttgggg agaacctgtt
      901 caaagaaatt gagcaaaacg aaattgagga gatcttttcc gttgacttaa gtgccatcac
      961 taatgatgga aataccgtta aacgctttga ctttgtgatc aaaaatgaac aggtcttata
     1021 cttgattgag gtcaatttct actctggaag tggctcgaag ttaaacgaaa ctgcacgctc
     1081 atacaaaatg attgccgaag agactaaggc tattccgaac gtggagttta tgtggatcac
     1141 tgacggacaa gggtggtaca aggccaaaaa caatttgcgc gagaccttcg acatcctgcc
     1201 tttcctttac aatattaacg atttggagca caatattctt aagaacctga aatgaagctt
     1261 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     1321 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     1381 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     1441 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1501 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     1561 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     1621 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     1681 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1741 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1801 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1861 tggtgaaagt aaaagatgct gaagatcagt tgggtgcagc aaactattaa ctggcgaact
     1921 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1981 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2041 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2101 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2161 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2221 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2281 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2341 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2401 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     2461 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2521 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     2581 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2641 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc
     2701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2941 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3001 cctacatacc tcgctctgct aatcctgtta ccagtcaggc atttgagaag cacacggtca
     3061 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     3121 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     3181 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     3241 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     3301 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     3361 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     3421 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     3481 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     3541 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     3601 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     3661 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     3721 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     3781 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     3841 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     3901 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     3961 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     4021 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     4081 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     4141 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     4201 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     4261 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     4321 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     4381 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     4441 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     4501 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     4561 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     4621 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     4681 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     4741 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     4801 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     4861 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     4921 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     4981 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     5041 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     5101 gataaaatat ttctaggctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     5161 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     5221 gtccggcgta gaggatctgc tcatgtttga cagcttatca tcgatgcata atgtgcctgt
     5281 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     5341 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     5401 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     5461 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     5521 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     5581 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     5641 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     5701 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     5761 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     5821 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     5881 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     5941 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     6001 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     6061 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     6121 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     6181 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     6241 atact
//
