LOCUS       sequence-62157-e        4909 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 4909)
  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 4909)
  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: pMHBAD14
FEATURES             Location/Qualifiers
     source          1..4909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             293..310
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(350..367)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      519..605
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      697..724
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        742..833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(834..902,903..1694)
                     /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(1052..1071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1736..2191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1823..1842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2033..2054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2302..2890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2791..2810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3044..3061
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3076..3216
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3142..3161)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3302..3324
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3463..3482
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3665..4543)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4570..4854
                     /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     4777..4796
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4881..4903
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatgatcaaa atagccacac gtaaatattt aggcaaacaa aatgtctatg acattggagt
       61 tgagcgcgac cataattttg cactcaaaaa tggcttcata gcttctaatt gtttcaatgg
      121 tacccagtac aaacttatcc tgaacggtaa aaccctgaaa ggtgaaacca ccaccgaagc
      181 tgttgacgct gctaccgcgg aaaaagtttt caaacagtac gctaacgaca acggtgttga
      241 cggtgaatgg acctacgacg acgctaccaa aacctacacc gtaacggaat tccatcatca
      301 tcatcatcat taagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
      361 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
      421 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
      481 tggggtcccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
      541 gcaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      601 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
      661 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
      721 tttttgcgtt tctacaaact ctttgtttat ttttctaaat acattcaaat atgtatccgc
      781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      841 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      901 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      961 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1021 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1081 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1141 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1201 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1261 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1321 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     1381 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1441 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1501 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1561 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1621 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1681 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttacgcg
     1741 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1801 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1861 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1921 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     1981 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2041 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     2101 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2161 aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga tcctttttga
     2221 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2281 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2341 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2401 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2461 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2521 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2581 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2641 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     2701 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2761 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2821 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2881 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2941 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3001 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3061 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3121 ccggatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     3181 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     3241 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3301 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcaa
     3361 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     3421 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     3481 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     3541 gatctaattc tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg
     3601 aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta
     3661 ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact
     3721 cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa
     3781 aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg
     3841 cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga
     3901 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
     3961 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
     4021 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
     4081 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
     4141 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
     4201 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
     4261 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
     4321 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
     4381 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
     4441 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa
     4501 acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc
     4561 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
     4621 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
     4681 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
     4741 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
     4801 gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt
     4861 ttttttgggc tagaaataat tttgtttaac tttaagaagg agatataca
//
