LOCUS       sequence-396440-        6599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of zebrafish TDP2 recombinant protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  TITLE     Oliver lab zebrafish expression plasmid
REFERENCE   2  (bases 1 to 6599)
  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: pAWO-SUMO-3C_14M_zTDP2
FEATURES             Location/Qualifiers
     source          1..6599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..35
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(80..127)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     123..141
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(994..1017)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(1312..1329)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1345..1367
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1382..1406
                     /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)."
     primer_bind     complement(1406..1425)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1407..1425)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1509..1528)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1734..1811
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1812..2894
                     /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     complement(1831..1850)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4206..4397
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4391..4413)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4499..4638
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4554..4573
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4653..4670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4824..5412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4904..4923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5583..6374,6375..6443))
                     /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     6206..6225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6444..6548)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattatccgg
       61 atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
      121 atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag
      181 cagccggatc tgctcgagcg gccgccagtg tgatggatat ctgcagaatt cttattaggt
      241 gttaaaggta caataaatac cccaatgatc gctggtatag cgaccacaat ccagtttttc
      301 cataccaatc agtgccatgt gatccggtgt cacaggcggt gcggtttttg cgctacgcag
      361 aaaaatacga tcaaaacgca gtttacatgc tgcggtaata cccagattgc tatttgcctg
      421 ggtatcccag gtataacgac aatgttcttg tttacccagc tgttcccaaa catcacaaac
      481 accggcaggc agaccaccaa catttgcaac ttcggcatca cgcagattgg tatcaccggc
      541 aaaaataaca atggcgtcct caggtgcttc tttgattttc tgcagcacaa cacgcagctg
      601 tttggtacgt tcttggctct gattacgggt actttccaga tggctggtca tcagatacag
      661 tttctgaccg ctaaaggtaa cctgtgcaat cagcagatta cgcatcatct gtgtggtcgg
      721 aaaacaaatg atttcgcttt ccagaaattt aacacggctt ttacgcagca taataccggt
      781 gaaataacca tcatcactac cttcaaaaaa caggtagcta actgcacgtt ttttcagata
      841 ctgcacatag gcaggaatca gttcttgcag aaaaacaaca tccggtgtat acagtgccag
      901 atagctacac agaccacgtg cacgatcgct cagattattg gtatccaggc catcaacatt
      961 ccagctaata atgctcagtt tgctatccat atggggcccc tggaacagaa cttccagacc
     1021 gccaatctgt tcacggtgag cttcgataat gtcgttatct tccatgtcca gatcttccgg
     1081 ggtttgatct gcctggatgc gaataccgtc atacaggaaa cgcagcgaat ccatttcttt
     1141 gccctgacgt ttagcaaatg cttccatcag gcgacgcagc ggcgtggttt tcttaatttt
     1201 aaagaaaatt tcagagctgc catctgagac tttcaggtta atatgggttt ccggtttgac
     1261 ttccggttta acttccggtt ttgcttcctg attgacttcg ctgtcgctgc cgtgatggtg
     1321 atggtgatgg ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt atttctagag
     1381 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
     1441 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     1501 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     1561 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     1621 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     1681 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     1741 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     1801 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     1861 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     1921 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     1981 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     2041 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     2101 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     2161 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     2221 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     2281 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     2341 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     2401 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     2461 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     2521 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     2581 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     2641 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     2701 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     2761 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     2821 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2881 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     2941 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     3001 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     3061 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     3121 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
     3181 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg
     3241 ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt
     3301 cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta
     3361 gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg
     3421 atcattggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg
     3481 ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt
     3541 gcatggagcc gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca
     3601 ccactccaag aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc
     3661 cttggcagaa catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg
     3721 gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc
     3781 taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt
     3841 gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt
     3901 tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga
     3961 tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc
     4021 gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt
     4081 taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca
     4141 tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg
     4201 catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga
     4261 cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg
     4321 aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg
     4381 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4441 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     4501 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
     4561 agagcagatt gtactgagag tgcaccattg cggtgtgaaa taccgcacag atgcgtaagg
     4621 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5221 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5881 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6181 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     6241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6421 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6481 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6541 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcat
//
