LOCUS       Exported                7957 bp ds-DNA     circular SYN 07-JUN-2012
DEFINITION  attB1-attB2.
ACCESSION   .
VERSION     .
KEYWORDS    Expression Clone/pET-DEST42/pD221-SSRP1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 15, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
COMMENT     Storage Type: Constructed
FEATURES             Location/Qualifiers
     source          1..7957
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        47..63
                     /label=T7 Promoter
     operon          66..90
                     /label=lacO
     misc_feature    132..152
                     /label=attB1
     CDS             158..2326
                     /label=SSRP1
     misc_feature    2083
                     /label=A2G
     misc_feature    2119
                     /label=C2T
     misc_feature    2199
                     /label=A2C
     misc_feature    2215
                     /label=C2G
     misc_feature    complement(2331..2351)
                     /label=attB2
     misc_feature    2338
                     /label=Gateway Cloning Description
                     /note="Generate PCR product using computed primers and a 
                     DNA polymerase (i.e. Platinum Pfx or Platinum Taq).	Check 
                     quality and purify product by PEG/Mg2+ precipitation and 
                     agarose gels.	Mix purified PCR product and pDONR Vector 
                     pDONR221 in BP buffer and TE buffers.	Thaw, vortex and add 
                     BP Clonase enzyme. Incubate >= 1 hr. at 25?.	Add Proteinase
                     K for 10 min at 37?.	Transform into DH5a or TOP10 cells and
                     plate on kanamycin plates.	Pick colonies and grow in 
                     selection media."
     misc_feature    2338
                     /label=Gateway Cloning Description
                     /note="Mix Entry Clone pD221-SSRP1 and Destination Vector 
                     pET-DEST42 in LR buffer and TE buffers	Thaw, vortex and add
                     LR Clonase enzyme (LR Clonase Plus enzyme for multi-site 
                     Gateway). Incubate >= 1 hr. at 25?.	Add Proteinase K for 10
                     min at 37?.	Transform into DH5a E. coli cells (One Shot 
                     TOP10 for multi-site Gateway) or any non-F' episome bearing
                     strain.	Select Expression Clones on agar plates with 
                     antibiotic ampicillin.	Express in appropriate host cells."
     misc_feature    2372..2413
                     /label=V5 epitope
     misc_feature    2423..2440
                     /label=6x his
     terminator      2455..2583
                     /label=T7 terminator
     CDS             2983..3843
                     /label=Amp(R)
     CDS             3988..4661
                     /label=pBR322 ori
     CDS             complement(5032..5223)
                     /label=ROP
ORIGIN
        1 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
       61 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
      121 aaggaatatc acaagtttgt acaaaaaagc aggcttaatg acagactctc tggagtacaa
      181 cgacataaac gccgaagtgc gcggagtctt gtgttccgga cgcctaaaga tgaccgagca
      241 gaacatcatc ttcaagaaca ccaagaccgg caaggtggag cagatctcgg cagaggacat
      301 agacctgatc aattcgcaga agttcgtggg cacctgggga ctgagggtgt tcaccaaagg
      361 cggcgtgctc caccgcttca ccggattccg cgacagcgag cacgagaagc tgggcaagtt
      421 tatcaaggct gcctactcgc aggagatggt cgagaaggag atgtgcgtca agggctggaa
      481 ctggggcacc gcccgcttca tgggctccgt cctgagcttc gacaaggagt cgaagaccat
      541 cttcgaggtg ccgctgtcgc acgtttcgca gtgcgtgacc ggcaagaacg aggtcaccct
      601 ggagttccac caaaacgacg atgcgcccgt gggtctactg gagatgcggt tccacatacc
      661 cgccgtggag tcggccgagg aggatccggt agacaagttc caccagaacg taatgagcaa
      721 ggcctcggtc atctcggctt cgggcgagtc catcgccatt ttcagagaga tccagatcct
      781 cacgcctcgc ggtcgctatg acatcaagat cttctcgacc ttcttccagc tgcacggcaa
      841 gacgttcgac tacaagattc ccatggactc ggtgctgcgg ctcttcatgc tgccccacaa
      901 agacagtcga cagatgttct ttgtgctctc cttggatccg cccatcaagc agggacaaac
      961 gcgttaccac tacctggtcc tgctgtttgc tcccgatgag gagaccacca ttgagctgcc
     1021 attctcggaa gccgagttgc gagacaagta cgagggcaag ctggagaaag agatctccgg
     1081 gccggtgtac gaggtgatgg gcaaagtgat gaaggtgctg atcggtcgaa aaattaccgg
     1141 acccggtaac tttatcggac actctggcac ggctgcagtg ggctgctcgt tcaaggctgc
     1201 agctggatat ctgtatcccc tggagcgagg attcatctat atccacaagc caccgctgca
     1261 tatccgcttt gaggagatta gttctgtgaa ctttgcccgc agcggcggat ccacgcgatc
     1321 tttcgacttc gaagtgacgc tcaagaacgg aactgttcac atcttctcct ccatcgagaa
     1381 ggaggagtat gccaagctct tcgactacat cacacagaag aagttgcatg tcagcaacat
     1441 gggcaaggac aagagcggct acaaggacgt ggactttggt gattcggaca acgagaacga
     1501 accagatgcc tatctggctc gcctcaaggc tgaggcgagg gaaaaggagg aggacgacga
     1561 cgatggcgac tcggatgaag agtccacgga tgaggacttc aagcccaacg agaacgagtc
     1621 cgatgtggcc gaggagtatg acagcaacgt ggagagtgat tcggacgatg acagcgatgc
     1681 tagtggcggc ggaggcgaca gcgacggcgc caagaaaaag aaggagaaga agtccgagaa
     1741 gaaagagaaa aaggagaaaa aacacaagga gaaggagaga acaaagaaac cctccaagaa
     1801 gaagaaggac tctggcaaac ccaagcgcgc caccaccgct ttcatgctct ggctgaacga
     1861 cacgcgcgag agcatcaaga gggaaaatcc gggcataaag gttaccgaga tcgccaagaa
     1921 gggcggcgag atgtggaagg agctgaagga caagtccaag tgggaggatg cggcggccaa
     1981 ggacaagcag cgctaccacg acgagatgcg caactacaag cctgaagcgg gcggtgacag
     2041 cgacaacgag aagggtggaa agtcctccaa gaagcgcaag acagagcctt ctccatccaa
     2101 gaaggcgaat acctcgggta gcggcttcaa gagcaaggag tacatttcgg acgacgactc
     2161 caccagctcc gacgacgaga aggacaacga gcctgccacg aagaagagca agccgccatc
     2221 cgacggcgat gccaagaaga aaaaggccaa gagcgagagc gaaccggagg agagcgagga
     2281 ggacagcaat gccagcgatg aggatgagga agatgaggcc agtgattacc cagctttctt
     2341 gtacaaagtg gtgatcaatt cgaagcttga aggtaagcct atccctaacc ctctcctcgg
     2401 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttgat ccggctgcta
     2461 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     2521 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     2581 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     2641 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     2701 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     2761 aacatgagaa ttaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2821 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2881 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2941 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3001 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3061 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3121 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3181 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3241 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3301 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3361 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3421 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3481 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     3541 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3601 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3661 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3721 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3781 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3841 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4141 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4681 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4741 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4801 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     4861 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4921 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4981 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5041 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     5101 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     5161 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     5221 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     5281 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     5341 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     5401 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     5461 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     5521 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     5581 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     5641 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
     5701 aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt
     5761 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct
     5821 tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg
     5881 ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc
     5941 catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt
     6001 ccagtgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg
     6061 tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa
     6121 gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg
     6181 tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     6241 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     6301 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     6361 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     6421 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     6481 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     6541 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6601 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     6661 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     6721 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     6781 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     6841 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     6901 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     6961 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     7021 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     7081 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     7141 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     7201 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     7261 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     7321 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     7381 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     7441 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     7501 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     7561 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     7621 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     7681 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     7741 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     7801 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     7861 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     7921 cgccagcaac cgcacctgtg gcgccggtga tgccggc
//
