LOCUS       sequence-429632-        6098 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses his-tagged T4 DNA ligase in E. coli hosts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6098)
  TITLE     Bacterial expression plasmids for production of recombinant PNK
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6098)
  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            "Unpublished"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-hisT4DNALig
FEATURES             Location/Qualifiers
     source          1..6098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(404..451)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     447..465
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1992..2009)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2020..2042
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2073..2092)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2074..2092)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2176..2195)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3653..3844
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3838..3860)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3946..4085
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4001..4020
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4100..4117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4271..4859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4351..4370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5030..5821,5822..5890))
                     /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     5653..5672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5891..5995)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6063..6081
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
      241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
      301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
      361 cacacccgtc ctgtggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
      421 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact
      481 cagcttcctt tcgggctttg ttagcagccg gatccgcata ggccttatag accagttacc
      541 tcatgaaaat caccaaatac atcttcgaat gtattagctt tagttttatc ttcacgtaaa
      601 cgaatcgcaa tcggaagaaa taatttaacg taatcagtgc ggccatcaga ttttaaccaa
      661 ccgttgcatt cgcactctag aatttttcca atataataat tttggttttc cataatgcga
      721 gtacggtcaa gttcatgcga ttttacaccg gctttatctt ttaagcctga accagcattt
      781 accttaattt ttccacactc tgactcaaga ataaatccac ccgctttagt agggtcttta
      841 cggtgaggat aaattcctac aatttttaaa tcaacatcaa ttacttcttt aaatttataa
      901 agattttttg aacgagcatt ttcccataat ccatcgatat ttttgagaat aataccttca
      961 agaccttggt caatatactt tttataaatt accttagctt catctaggtt atttactacc
     1021 tggttttcaa ttaaaattac tttatcatat ccagatgtca tttgttctag tttagaaaaa
     1081 cgtacatcat atttcaaacg aaatgcagga agactgtata tttctaccaa cgggacataa
     1141 tcccagacct gaaacttcat gcattgtgct tctttttcag aaatggttcc ctttaaagat
     1201 ttattggcga ttccattaga agcagtacgt gattcagcaa cttcggcgaa ttctttagct
     1261 ttactgtttt caggataagc atcaaaaaga aaatctaggc cttctggctc ctttttaact
     1321 tgctcatggt ataccaattc gccatcaatc aacacacctt ctggatgaat ctggcgggct
     1381 tcagcggtca ttttaattaa ctcttcctta agaagatcta atcctagata ttcattacca
     1441 gctcgtgata aaagacgaac atcatctaat tcatcacctc taacttcagc aaaacaccga
     1501 gctccatcag cttttaactg agcaaaggct ggaaatttga tattcttatt aatgcctttt
     1561 tcatcataag aacttgcgag catttgaggt tgttcaggaa ttaaacctgg ccaaactttg
     1621 tttgcaatag atactgaagc accacattca aggtctcgca tcatcactcg acgcaaaact
     1681 tcaacatcat cttttttacc atcggtgata tatccagtta attcctcaat tgctgcattt
     1741 ccagtcaatt tccgagtagc taatgtgaat tcaatgaagt caagcatatc ggtaagagtc
     1801 aacattccaa aactctgggt agcaatacca ggtttaggcc atttcttgat ataatactgt
     1861 aacccacgag aataagtcag acgatatact cgtttaagca attcattatc tttattcttt
     1921 tcaagaattg cttgcttctg tttagttgaa ccaatagatg ctatttcgtt cagaatttta
     1981 agaatgccgc tgtgatggtg atggtggtgc atatgtatat ctccttctta aagttaaaca
     2041 aaattatttc tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg
     2101 ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     2161 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     2221 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     2281 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
     2341 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc
     2401 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     2461 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2521 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     2581 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     2641 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     2701 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     2761 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     2821 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc
     2881 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     2941 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     3001 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     3061 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     3121 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     3181 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     3241 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     3301 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     3361 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     3421 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     3481 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     3541 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     3601 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     3661 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     3721 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     3781 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     3841 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3901 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     3961 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     4021 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4081 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4141 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4201 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4261 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4321 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4381 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4441 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4501 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4561 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4621 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4681 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4741 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4801 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4861 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4921 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4981 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5101 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5161 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5221 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5281 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5341 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5401 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5461 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5521 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5581 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5641 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5701 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5761 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5821 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5881 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5941 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6001 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6061 aaaataggcg tatcacgagg ccctttcgtc ttcaagaa
//
