LOCUS       sequence-466923-        5134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged Beckwith-Wiedemann 
            syndrome chromosomal region 1 candidate gene B protein LCD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5134)
  TITLE     Mittag bacterial expression plasmids
REFERENCE   2  (bases 1 to 5134)
  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"
COMMENT     Sequence Label: pDest17_Beckwith-Wiedemann syndrome chromosoma...
FEATURES             Location/Qualifiers
     source          1..5134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             167..187
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(603..627)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(680..698)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      694..741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1107..1112,1113..1129,1130..1135))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1162..1180)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1248..1352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1353..1421,1422..2213)
                     /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(1571..1590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2384..2972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2873..2892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3126..3143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3158..3297
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3223..3242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3383..3405
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3399..3590)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5052..5071
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggctccgaaa acctgtattt
      181 ccagggtagc atgggtgaac tgccgggtag cgaaggtatg tgggaaaatt gcccgctggg
      241 ttgggttaaa aagaaagcaa gcggtaccct ggcaccgctg gattttctgc tgcagcgtaa
      301 acgtctgtgg ctgtgggcaa gcgaaccggt tcgtccgcag ccgcagggta ttcatcgctt
      361 tcgtgaagcc cgtcgtcagt tttgccgtat gcgcggtagt cgtctgaccg gtggccgtaa
      421 aggctttggc agtagtggtc tgcgttttgg tcgtggtggt tttagtgaag aagtgatgcc
      481 gcagccggtg ctgaaagcaa tgcgttgtgc agaaggtgca tggtggttta gcccggatgg
      541 tccggcaggt agtgcagcca gcatttggcc ggcagaaggc gccgaaggcc tgcctggtta
      601 aaacccagct ttcttgtaca aagtggttga ttcgaggctg ctaacaaagc ccgaaaggaa
      661 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      721 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc acaggacggg
      781 tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga gcaggactgg
      841 gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa attgcatcaa
      901 cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat ggacgatatc
      961 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     1021 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     1081 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     1141 agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1201 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1261 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1321 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1381 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1441 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1501 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1561 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1621 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1681 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1741 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1801 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1861 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     1921 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1981 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2041 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2101 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2161 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2221 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2281 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2341 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2401 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2461 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2521 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2581 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2641 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2701 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2761 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2821 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2881 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2941 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3001 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3061 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3121 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3181 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     3241 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3301 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3361 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3421 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     3481 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     3541 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     3601 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     3661 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     3721 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     3781 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     3841 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     3901 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     3961 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     4021 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     4081 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     4141 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     4201 ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc
     4261 gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt
     4321 tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag
     4381 ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct
     4441 gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca
     4501 taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt
     4561 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     4621 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     4681 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     4741 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     4801 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgatcgacgc
     4861 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     4921 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     4981 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     5041 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     5101 atgccggcca cgatgcgtcc ggcgtagagg atcg
//
