LOCUS       sequence-429684-        5781 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses his-tagged N-terminus truncated TdT in E. coli hosts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5781)
  TITLE     Bacterial expression plasmids for production of recombinant PNK
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5781)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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-hisTdT
FEATURES             Location/Qualifiers
     source          1..5781
                     /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"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             84..113
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1311..1329)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1325..1372
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1738..1743,1744..1760,1761..1766))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1793..1811)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1879..1983
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1984..2052,2053..2844)
                     /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(2202..2221)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3015..3603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3504..3523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3757..3774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3789..3928
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3854..3873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4014..4036
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4030..4221)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5679..5698
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcaccatc atcatcatca tcaccaccac catatgcgta
      121 ccgattacag cgcgaccccg aacccgggct ttcagaaaac cccgccgctg gcggtgaaaa
      181 aaatcagcca gtacgcgtgt cagcgcaaaa ccaccctgaa caactataac cacattttta
      241 ccgatgcgtt tgaaattctg gcggaaaaca gcgaatttaa agaaaacgaa gtgagctatg
      301 tgacctttat gcgcgcggcc agcgtgctga aaagcctgcc gtttaccatt atctccatga
      361 aagataccga aggcatcccg tgtctgggcg ataaagtgaa atgtattatt gaagaaatta
      421 ttgaagatgg cgaatcatca gaagtgaaag ccgtgctgaa cgatgaacgc tatcagagct
      481 ttaaattatt taccagcgtg tttggcgtgg gtctgaaaac ctcagaaaaa tggtttcgca
      541 tgggttttcg cagcctgtcc aaaattatgt ctgacaaaac gctgaaattt acgaaaatgc
      601 agaaagctgg ttttctgtat tatgaagacc tggtatcgtg tgtaacccgc gctgaagctg
      661 aagccgtagg tgtactcgta aaagaagctg tttgggcttt tctgccggat gcctttgtta
      721 caatgacagg tggctttcgc cgtggcaaaa aaatcggcca cgatgttgat tttctgatca
      781 cctccccggg ttccgccgaa gatgaagaac agcttttacc gaaagtgatc aatctctggg
      841 aaaaaaaagg tctgctgctc tattatgatc tagtggaatc gacctttgaa aaattcaaac
      901 tcccgagtcg gcaggtggat acgctggatc atttccagaa atgtttcttg atcctgaaac
      961 tacatcatca gcgtgtggat agtagcaaaa gcaatcagca ggaaggtaaa acatggaaag
     1021 ccatccgtgt tgatttggtg atgtgtccgt atgaaaatcg tgccttcgcc ctgctgggct
     1081 ggaccggctc ccgtcagttc gaacgtgata ttcgtcgcta tgcgacccat gaacgtaaaa
     1141 tgatgctgga taatcatgcg ctgtatgata aaaccaaacg tgtgtttctg aaagcggaaa
     1201 gcgaagaaga aatttttgcg catctgggcc tggattatat tgaaccgtgg gaacgtaatg
     1261 cgtaacggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1321 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1381 aggaactata tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag
     1441 tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc
     1501 cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt
     1561 gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa
     1621 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
     1681 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
     1741 aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg
     1801 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     1861 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1921 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1981 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2041 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2101 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2161 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2221 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2281 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2341 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2401 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2461 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2521 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2581 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2641 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2701 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2761 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2821 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2881 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2941 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3001 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3061 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3121 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3721 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3781 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     3841 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3901 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3961 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4021 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     4081 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     4141 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     4201 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     4261 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     4321 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     4381 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     4441 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     4501 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     4561 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     4621 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     4681 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     4741 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     4801 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     4861 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     4921 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     4981 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     5041 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     5101 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     5161 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     5221 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     5281 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5341 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     5401 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5461 aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat taggaagcag
     5521 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     5581 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     5641 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     5701 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     5761 gagatctcga tcccgcgaaa t
//
