LOCUS       sequence-345439-        5992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses His-tagged uracil-DNA-glycosylase enzyme from marine 
            bacterium BMTU 3346 for the purpose of cross contamination 
            prevention in nucleic acid amplification reactions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5992)
  TITLE     Robust and inexpensive molecular diagnostic s with lyophilized open 
            access RT - LAMP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5992)
  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            "Unpublished"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: pET-21a(+)-BMTU UDG-6xHis
FEATURES             Location/Qualifiers
     source          1..5992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             69..91
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             102..119
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      808..855
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      892..1347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(979..998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1189..1210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1373..1477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1478..1546,1547..2338)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1696..1715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2509..3097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2998..3017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3251..3268
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3283..3422
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3348..3367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3508..3530
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3524..3715)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4524..5606)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5568..5587
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5607..5684)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5890..5909
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta ggatcatgca
       61 taaataattt tgtttaactt taagaaggag atatacatat gcatcatcat caccaccatg
      121 agctgatttc accgcttgac cctggatggg acgccgcgtt ggctgcgcag acagaggcct
      181 tggatgttgt tggaaaagac ctggttcgtc gtcgcgtagc cggtgaatat atcctgcctg
      241 ctccagagca cgtgttacgt tcttttcgcc aacctttcga tcaagtcaag gtgttggtat
      301 taggacaaga cccgtaccca acacctggac acccaatcgg attgagcttc gcagtggatc
      361 gtgacgtccg ccctttacca cgcagtcttg tgaacattta tcaagaattg tccactgact
      421 tggggattcc acctgctagt catggggacc tgacggcttg gaccgagcaa ggggttctta
      481 tgctgaaccg cgtcttgact gttcgtgccg gagcagcggc atctcaccgc ggcatcggtt
      541 gggagcaaat tacacaaacc gctgtggaag cccttgccgc acgtggtaca cccctggtag
      601 ccattttatg gggcaacgat gcccgcaaaa tggctcccgt gcttcgccaa ggcggagcaa
      661 cggcaattat tgagtctccg catccctcac ccttatccgc tcatcgtggc ttctttggga
      721 gtcgtccatt ctcgcgtaca aacgagatgc ttgagcagtt gggtgcagat cctgtcgatt
      781 ggcgtgttga acgtgactga gcaataacta gcataacccc ttggggcctc taaacgggtc
      841 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
      901 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      961 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1021 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1081 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1141 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1201 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1261 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1321 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     1381 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1441 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1501 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1561 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1621 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1681 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1741 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1801 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1861 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1921 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1981 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     2041 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2101 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2161 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2221 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2281 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2341 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2401 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2461 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2641 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3121 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3181 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3241 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     3301 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     3361 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3421 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3481 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3541 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3601 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3661 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3721 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3781 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3841 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3901 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3961 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     4021 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     4081 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     4141 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     4201 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4261 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4321 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4381 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4441 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4501 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4561 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4621 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4681 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4741 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4801 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4861 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4921 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4981 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     5041 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     5101 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     5161 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     5221 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     5281 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     5341 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     5401 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     5461 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5521 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5581 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5641 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5701 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5761 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5821 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5881 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5941 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at
//
