LOCUS       sequence-468719-        5605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminally 6His tagged THRAP3_THSD7A 
            (THRAP3) fusion protein LCD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5605)
  TITLE     Mittag bacterial expression plasmids
REFERENCE   2  (bases 1 to 5605)
  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_THRAP3_THSD7A fusion protein LCD (663-...
FEATURES             Location/Qualifiers
     source          1..5605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..39
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(66..71,72..88,89..94)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(460..507)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     503..521
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    574..598
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1485..1505)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1508..1532)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1542..1559)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1579..1601
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1632..1651)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1633..1651)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1735..1754)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3216..3407
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3401..3423)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3509..3648
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3564..3583
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3663..3680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3834..4422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3914..3933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4593..5384,5385..5453))
                     /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     5216..5235
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5454..5558)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct
       61 catgtttgac agcttatcat cgataagctt taatgcggta gtttatcaca gttaaattgc
      121 taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc tcatcgtcat cctcggcacc
      181 gtcaccctgg atgctgtagg cataggcttg gttatgccgg tactgccggg cctcttgcgg
      241 gatatcgtcc attccgacag catcgccagt cactatggcg tgctgctagc gctatatgcg
      301 ttgatgcaat ttctatgcgc acccgttctc ggagcactgt ccgaccgctt tggccgccgc
      361 ccagtcctgc tcgcttcgct acttggagcc actatcgact acgcgatcat ggcgaccaca
      421 cccgtcctgt ggatatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg
      481 tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc
      541 ttcctttcgg gctttgttag cagcctcgaa tcaaccactt tgtacaagaa agctgggttt
      601 tattcggtgg tcggctgaat attatctttc tcttcacgat tttcggtacc actttcatca
      661 tcttcaattt cgccttcttc accagaaaat ttatcatggg cccatttcgg gctggtgcta
      721 cttttacgaa acataaagcg tccgcgacca cgcggaaatg caccgcggcc acggccacgg
      781 ctaacccatt tatcactacc ttcgccttca cggtcatcat gcaggtaata ttttttgctt
      841 ttcggggtat attccggatc ccattcttct tcacgattac gtttctggaa atcgttgttg
      901 ctgttgttat tattattacc actataatta ccgcggcccc aaccgcgacc acgtgcgcga
      961 aactgaaagg taccacgtgc acgaccgccg ccacgttcgc tcggacccac aaaatctttg
     1021 gtacccaggc ggcttttatc aaaaccggtg gtactttctt cgcgttcttc ggtttcttcg
     1081 gtatattctt ctgctttata gctatgacta ctctggctac tactgctgct actgcggcta
     1141 cgatcgcgtg cgcggcggtg ttttttctgt tttttgctac ctttatgggt tttttcggtt
     1201 ttttcggcgc tgcgttcgcg gctatggctg ctatcacggc tatcaacgct ttcacggctt
     1261 ttgccacgtt tcagatcgcg ctctttatgt tttttacgac gttcaatatc cagacgcaga
     1321 tccaccggat catctttata tttgccttcg gctttataac ccggttcacg cggacttttc
     1381 atttcatcat gggccagacc atgtttacga aaggtactcg ggctaatatc aatacgacga
     1441 tgaatttccg gacttttctt attttttgcg gcttcctgtt cgctgccctg gaaatacagg
     1501 ttttcggagc ctgctttttt gtacaaactt gttgattcga ggtgatggtg atggtgatgg
     1561 tagtacgaca tatgtatatc tccttcttaa agttaaacaa aattatttct agagggaaac
     1621 cgttgtggtc tccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct
     1681 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
     1741 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt
     1801 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc
     1861 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
     1921 taatgcagga gtcgcataag ggagagcgtc gatcgaccga tgcccttgag agccttcaac
     1981 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     2041 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     2101 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     2161 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     2221 gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg
     2281 cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc
     2341 gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga
     2401 tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt cacggcgatt
     2461 tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac
     2521 cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg
     2581 gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct
     2641 tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca
     2701 tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca
     2761 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
     2821 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
     2881 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
     2941 agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     3001 gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
     3061 ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
     3121 gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
     3181 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
     3241 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     3301 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     3361 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3421 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3481 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     3541 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg
     3601 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     3661 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3721 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3781 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3841 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3901 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3961 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4021 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4081 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4141 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4201 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4261 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4321 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4381 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4441 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4501 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4561 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4621 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4681 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4741 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4801 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4861 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     4921 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4981 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5041 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5101 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5161 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     5221 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5281 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5341 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5401 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5461 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5521 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5581 cctgacgtct aagaaaccat tatta
//
