LOCUS       Exported                6639 bp ds-DNA     circular SYN 29-AUG-2024
DEFINITION  Ligation of L into v.
ACCESSION   .
VERSION     .
KEYWORDS    pET28.His.3C.86b.8D2Q.Tau2N4R
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6639)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 29, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Vector molecule:  v, pET28.His.3C.hTau2N4R.LMW
               Fragment ends: BgLII and BsiW1
               Fragment size: 6399, 207
            Insert molecule:  fragment BgLII.his.3C.86b.Tau.BsiW1
              Fragment size:  300
            maxi prep is confiremd by Rapid Plasmid Sequencing#229926n.RPS
            maxi is confirmed by Rapid plasmid Sequencing#244528 (used for 
            recombinant protein production)
FEATURES             Location/Qualifiers
     source          1..6639
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             472..1881
                     /codon_start=1
                     /label=ORF
                     /translation="MEHHHHHHLEVLFQGPVSGWRLFKKISHMAEPRQEFEVMEDHAGT
                     YGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVT
                     APLVDEGAPGKQAAAQPHTEIPEGTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGT
                     GSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKDPPSSGEPPKSGDR
                     SGYDDPGDPGTPGSRSRTPSLPDPPTREPKKVAVVRDPPKSPSSAKSRLQTAPVPMPDL
                     KNVKSKIGSTENLKHQPGGGKVQIINKQLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKP
                     VDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSQIGSLDNITHVPGGGNKKI
                     ETHKLTFRENAKAKTDHGAEIVYKSPVVSGDDDPRHLSNVSSTGSIDMVDSPQLATLAD
                     EVSASLAKQGL"
     CDS             478..495
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             496..519
                     /product="recognition and cleavage site for human  
                     rhinovirus 3C and PreScission proteases"
                     
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human  rhinovirus 
                     3C and PreScission proteases"
     misc_feature    520..552
                     /label=86b
     misc_feature    559..1879
                     /label=LMW2N4R
     CDS             1888..1905
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(2603..3418)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /note="Kanr"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    4314..4453
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="basis of mobility region from pBR322"
     CDS             complement(4555..4746)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy 
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5555..6637)
                     /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)."
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
       61 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
      121 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
      181 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
      241 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
      301 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
      361 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
      421 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac catggaacat
      481 catcatcatc atcatctgga ggtgctgttc caaggtcctg tgagtggctg gcgactgttc
      541 aagaagatca gccatatggc tgagccccgc caggagttcg aagtgatgga agatcacgct
      601 gggacgtacg ggttggggga caggaaagat caggggggct acaccatgca ccaagaccaa
      661 gagggtgaca cggacgctgg cctgaaagaa tctcccctgc agacccccac tgaggacgga
      721 tctgaggaac cgggctctga aacctctgat gctaagagca ctccaacagc ggaagatgtg
      781 acagcaccct tagtggatga gggagctccc ggcaagcagg ctgccgcgca gccccacacg
      841 gagatcccag aaggaaccac agctgaagaa gcaggcattg gagacacccc cagcctggaa
      901 gacgaagctg ctggtcacgt gacccaagct cgcatggtca gtaaaagcaa agacgggact
      961 ggaagcgatg acaaaaaagc caagggggct gatggtaaaa cgaagatcgc cacaccgcgg
     1021 ggagcagccc ctccaggcca gaagggccag gccaacgcca ccaggattcc agcaaaaacc
     1081 ccgcccgctc caaaggatcc acccagctct ggtgaacctc caaaatcagg ggatcgcagc
     1141 ggctacgatg atcccggcga tccaggcact cccggcagcc gctcccgcac cccgtccctt
     1201 ccagatccac ccacccggga gcccaagaag gtggcagtgg tccgtgatcc acccaagtcg
     1261 ccgtcttccg ccaagagccg cctgcagaca gcccccgtgc ccatgccaga cctgaagaat
     1321 gtcaagtcca agatcggctc cactgagaac ctgaagcacc agccgggagg cgggaaggtg
     1381 cagataatta ataagcagct ggatcttagc aacgtccagt ccaagtgtgg ctcaaaggat
     1441 aatatcaaac acgtcccggg aggcggcagt gtgcaaatag tctacaaacc agttgacctg
     1501 agcaaggtga cctccaagtg tggctcatta ggcaacatcc atcataaacc aggaggtggc
     1561 caggtggaag taaaatctga gaagcttgac ttcaaggaca gagtccagtc gcagattggg
     1621 tccctggaca atatcaccca cgtccctggc ggaggaaata aaaagattga aacccacaag
     1681 ctgaccttcc gcgagaacgc caaagccaag acagaccacg gggcggagat cgtgtacaag
     1741 tcgccagtgg tgtctgggga cgatgatcca cggcatctca gcaatgtctc ctccaccggc
     1801 agcatcgaca tggtagactc gccccagctc gccacgctag ctgacgaggt gtctgcctcc
     1861 ctggccaagc agggtttgta gctcgagcac caccaccacc accactgaga tccggctgct
     1921 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1981 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     2041 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2101 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2161 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2221 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2281 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2341 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2401 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2461 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2521 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2581 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     2641 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     2701 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     2761 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     2821 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     2881 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     2941 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     3001 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     3061 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     3121 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     3181 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     3241 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     3301 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     3361 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     3421 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     3481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3661 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3901 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4201 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4261 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4321 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     4381 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4441 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4501 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4561 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4621 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4681 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4741 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     4801 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     4861 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     4921 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     4981 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     5041 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     5101 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     5161 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     5221 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     5281 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     5341 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     5401 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     5461 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     5521 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5581 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5641 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     5701 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     5761 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     5821 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     5881 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     5941 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     6001 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     6061 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     6121 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     6181 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     6241 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     6301 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     6361 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     6421 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     6481 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     6541 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     6601 catactctgc gacatcgtat aacgttactg gtttcacat
//
