LOCUS       Exported                8858 bp ds-DNA     circular SYN 16-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 17, 2020 from SnapGene Viewer 5.0.6
            https://www.snapgene.com
COMMENT     M13F failed
FEATURES             Location/Qualifiers
     source          1..8858
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..307
                     /locus_tag="F1 Ori"
                     /label=F1 Ori
     misc_feature    326..394
                     /locus_tag="lacz a"
                     /label=lacz a
     misc_feature    465..482
                     /locus_tag="M13F"
                     /label=M13F
     misc_feature    524..543
                     /locus_tag="5' homology fwd "
                     /label=5' homology fwd
     misc_feature    535..543
                     /locus_tag="NotI"
                     /label=NotI
     misc_feature    543
                     /locus_tag="XbaI"
                     /label=XbaI
     misc_feature    544..564
                     /locus_tag="F primer"
                     /label=F primer
     misc_feature    544..561
                     /locus_tag="Beginning of next 5' gene(1)"
                     /label=Beginning of next 5' gene
     misc_feature    562..569
                     /locus_tag="Beginning of next 5' gene"
                     /label=Beginning of next 5' gene
     misc_feature    1168..1187
                     /locus_tag="hrp-1_pro_1F"
                     /label=hrp-1_pro_1F
     misc_feature    1909..1929
                     /locus_tag="hrp-1_pro_2F"
                     /label=hrp-1_pro_2F
     misc_feature    2656..2678
                     /locus_tag="hrp-1_pro_3F"
                     /label=hrp-1_pro_3F
     misc_feature    3383..3409
                     /locus_tag="hrp-1_pro_4F"
                     /label=hrp-1_pro_4F
     misc_feature    3830..3856
                     /locus_tag="Seq_primer_stop_exon1/2 B9 CRISPR box"
                     /label=Seq_primer_stop_exon1/2 B9 CRISPR box
     misc_feature    3920..4136
                     /locus_tag="5' UTR"
                     /label=5' UTR
     misc_feature    3921..3940
                     /locus_tag="hrp-1_5'_1F"
                     /label=hrp-1_5'_1F
     misc_feature    4059..4085
                     /locus_tag="Exon1"
                     /label=Exon1
     misc_feature    4130..4136
                     /locus_tag="SanDIII/PpumI"
                     /label=SanDIII/PpumI
     misc_feature    4137..4955
                     /locus_tag="5' Homology arm"
                     /label=5' Homology arm
     misc_feature    4166..4901
                     /locus_tag="Exon2"
                     /label=Exon2
     misc_feature    4203..4208
                     /locus_tag="BstbI"
                     /label=BstbI
     misc_feature    4205..4206
                     /locus_tag="ins C between in 5'F and pro4F and HsR"
                     /label=ins C between in 5'F and pro4F and HsR
     misc_feature    4624..4643
                     /locus_tag="seqprimer1"
                     /label=seqprimer1
     misc_feature    4771..4776
                     /locus_tag="NruI"
                     /label=NruI
     misc_feature    4915..4934
                     /locus_tag="repl_seq_1F"
                     /label=repl_seq_1F
     misc_feature    4929..4955
                     /locus_tag="5' homology rev"
                     /label=5' homology rev
     misc_feature    4956..5231
                     /locus_tag="optimized HS LC"
                     /label=optimized HS LC
     misc_feature    5076..5078
                     /locus_tag="D to V"
                     /label=D to V
     misc_feature    5077
                     /locus_tag="AtoT (HS mutation, most used Val codon)"
                     /label=AtoT (HS mutation, most used Val codon)
     misc_feature    5232..6451
                     /locus_tag="3' Homology arm"
                     /label=3' Homology arm
     misc_feature    5232..5262
                     /locus_tag="3' homology fwd"
                     /label=3' homology fwd
     misc_feature    5248..5269
                     /locus_tag="hrp-1_repl_seq_R"
                     /label=hrp-1_repl_seq_R
     misc_feature    5427..5446
                     /locus_tag="seqprimer2"
                     /label=seqprimer2
     misc_feature    6420..6478
                     /locus_tag="3' homology rev"
                     /label=3' homology rev
     misc_feature    6457
                     /locus_tag="BamHI"
                     /label=BamHI
     misc_feature    6571..6591
                     /locus_tag="M13R"
                     /label=M13R
     misc_feature    6597..6619
                     /locus_tag="LacO"
                     /label=LacO
     misc_feature    6959..7641
                     /locus_tag="ColE1 Ori"
                     /label=ColE1 Ori
     misc_feature    7739..8398
                     /locus_tag="AmpR"
                     /label=AmpR
ORIGIN
        1 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
       61 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      121 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      181 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      241 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      301 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
      361 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      421 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      481 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc ggtggcggcc
      541 gctgtaggag ctgagcccat cttcaatgcc tccgattttc caaaattaaa cactttttcc
      601 atgaattctt ccattaattt ggcaattgag ccaaatgaaa agccagaaga gccagatgga
      661 gccggtgttg gagttggagc agacagagtt gaacggagtt ctggagcagc tggggttggg
      721 gtcggagcag caagagttga acgtggttca gggtcggcga ggaactgaaa atctaggttt
      781 tttaggaaaa attcgcctaa aaacatgccg aaaaaatttc cgaattttag acaattctgg
      841 aaaaaattgt agtgaaattg tctaaaaatt ttggtaaaag tttgcagaaa actgtcagaa
      901 aatgttaatt tcttacgtgt ttccttgctt gggacactag aacagcaccg aataagacaa
      961 aaagtacaaa acacagcttc atgttggaat ttatgccaga aaaatagctg taactcggct
     1021 tttatagaat agaattggtt cctggtgatg ctggagttct attaaacatg agaacaggag
     1081 tatgaaccgt gggaagtggt gttttggttg gagaggtgtt ctgcacggct cggcaattgt
     1141 ggtgcatcag tgtccaccaa acatcaagca taacctggat gataaggaga caagctttgg
     1201 tgcaaaagtt ttcaaatttt aaagcaattt ttaggactct tcaaagcatg aaactgtttc
     1261 tcaaggtttc cttctacttc taactccttc tagatgcatc acggcctcaa aatgtaaaac
     1321 ctcagaaatg gtaccagagg ttcaaaaatg caaatatcag ggacctgggc tagatcttct
     1381 cgtttcgagg gctgggaaac tgcccttccg tgtgaaaggg gcaattgtca ggcctggagc
     1441 atgatattga tcggttttgt gagaagacca acaaggtagc taataaatcg atcctatatt
     1501 gtattaacac atgtatttga atcagatctt tcatttctca ttgactccct caaatttcca
     1561 gcaagaatgg atgctacaat tgtaagttta accacaaatc atttttaatt atacctgaat
     1621 ttcttttcag accaaaacca gaaaagtcgt attcatatag tgttgttcaa gagattgtca
     1681 gttgtatgtg cctaacaaaa aaaaatctat ggcattctaa atttacaatt tcagcatagt
     1741 cgctcgccat gtgtcaccac tctttcgcca aaccatcgat ttgatggatc caggcattga
     1801 gaaaattcat ttactcagtt ctgccagtcc gtagtgtagt ttttaccgga tatagagaat
     1861 tgaatctaga aaacctggct gcctggcaag aagtcactgg tagtaacagg ctgatgaggg
     1921 cagccttcgg tgagctatcc gacgtgatgg gcatgccgaa acttagacta agaaagttca
     1981 gttttgataa cacatatgga ctggttgggt tatctgggaa agagaatatg caatctgtgg
     2041 atgactgctt caaggctatg aacttaccac atgcgaaggt gcttgaagtg actggaatcg
     2101 actttgtgga caatatttcg atggtgaagt actttcgccc aggttatctc gagaaaatcc
     2161 gcatgtattc ttgttattgc tcagctagcc aggagatgag tgaagaaatt gcaaatctcg
     2221 agcagtggaa gcaagcgaaa cattttctgt gcattggtgg tagaatgaac attcctattc
     2281 aaagattttt ccactttgac cagttctcca tcggtcttca gactctatca atcgatgatg
     2341 ctgttcagat aagagatgtg agttggtagt taaagtagcg ttttgtaatg tattctatat
     2401 tccagattct cctcagatcg cccagcttcg agtcagcaac tatcgatgtt caactacttg
     2461 aaaaagaagc cattgagaac gtctttacgc cgaatcacct attaatttat agaggaagag
     2521 agttcagacc ggtgggaact gattccagtt ttgtgatcaa tataacaaaa aacaagcttg
     2581 aaatccacaa gcacaaggga gctaatccac cacgtcgtag agacgaacgg ccaatccacg
     2641 agcgtatacg tcaaaggcaa gatggatggt gaagtttcaa tggttgctct aatacgttta
     2701 catgtctatc aaaccttttt ctgttactta agtaaatcgt tgtttcatat ttaaccagtc
     2761 ttacctacgg ctggcatatc ggacgtccgg cgtcagaacc ttgtgcgtct ggttttagat
     2821 gtttgaaaat tttggaaaat tattaaagtc tattgataac ctgtcaaaac cggacgaccg
     2881 gacgccaaat atgacagccc tagtcttacc agtataatat ttttataaac ctaaaacagg
     2941 tatgaatatc tcttttttct gcgcagcgct gggatgcttc tgtttcagaa aaatcacatg
     3001 gtataaaaaa tttccaaatt acagtagtcc taccgtaatc ctgcattatc ccactactgt
     3061 atgtaccttg acattatttt tcctcgccca actcccagcc caatatatct cttcaacagc
     3121 taagaactac atttttccta aactacagta aaccaacagt atccctacag taccacatta
     3181 attataaatt aaaacattat ttttcataag aaatcgtgag aatttcgaga aaaataaagg
     3241 attcaaagat tatttgaacg tttttttcgt gaaaaaatgt caaagaaatc aaaatgttca
     3301 tcttccttgt gaaaaatccg aaaagccatt ataaactagt tgattttcat aaaaatttgt
     3361 agaaatattt ttcttaaaat ttgtagaaga attttaacgt tttttaaaca aaaattccaa
     3421 aattggaatt tggtattgaa aatacttttt aaaagttatt tttttaattt gtaaaaattt
     3481 aactaaattc aaaaaataaa gtttccctcc tttttttctt aaaggcacac acatttatcg
     3541 actcgggtct cgccgcgaac agacgcgacg cgacgcggtt aggaatgcgg cgcgtcgcgt
     3601 gtaacccttt tctctgtggt tttcgagagc attctctctc ggttttcgtc acaaaaaggt
     3661 atgttttaca ggttttccgc gcactgtttt cgcgtaattc ttcattttaa ttcagataat
     3721 ctaattgttg tgtagtttta gctcgaattt atccgtaatc tttgattaaa tacgaataaa
     3781 actcgctaaa atagtcttat ttctcagata tttatcgatt tgttattttc cacgaatagt
     3841 ctactcgtgc atggcgtcta tatttaactt aataatctat ttttagattt atctttactc
     3901 taacttgtac ctatttcagt gatcaacacc tctgtcactc tttgtgagta atagcgttcg
     3961 ataggtagtt acaccgtgaa actagcctgt ctatttccta cttcataagc taagaactac
     4021 ttgttagcac aatccgatcg actactaaga actagatcat gacggacgtg gaaatcaagg
     4081 ctgaggttac gtttgttcac ttgcttttag ggtgtaggcc acctgatttg ggtcccgatc
     4141 atttaattat cgattttatt ttcagaatgg aagcggagat gcctcgctcg agccagagaa
     4201 ccttcgaaag attttcgttg gaggactcac gtcaaacacc accgatgacc tgatgcgaga
     4261 gttctactca caattcggag aaatcactga tatcatcgtc atgagagatc caactaccaa
     4321 gagatcccgc ggattcggat ttgtcacctt ctctggcaaa actgaagtcg atgctgccat
     4381 gaaacaacgc ccgcacatca tcgacggaaa gaccgtggac ccgaagcgtg ccgtgccacg
     4441 tgatgataag aaccgctccg agtcgaatgt ctccaccaaa cgcctgtacg tcagcggagt
     4501 tcgtgaggat cacaccgagg acatgctcac cgagtatttc acaaagtatg gaaccgtcac
     4561 caaatccgag attattctcg acaaagccac ccaaaagcca agaggcttcg gattcgtcac
     4621 cttcgatgat catgactctg tggatcaatg cgttcttcaa aaatcccata tggtcaacgg
     4681 acacagatgt gacgtgcgca aaggactctc caaggacgag atgagcaagg ctcagatgaa
     4741 ccgcgatcgt gaaacccgtg gcggaagatc tcgcgatgga cagcgtggcg gttacaatgg
     4801 cggtggtggt ggcggcggcg gctggggagg accagctcaa cgcggcggtc caggagcata
     4861 tggtggacca ggaggtggcg gccaaggagg atatggcgga ggtagttgga ttctggcttg
     4921 tgaattctcg attctaatat tattaacttt cagattacgg aggaggacgc ggtggttatg
     4981 gtggtggagg accaggatat ggaaatcagg gtggaggata tggaggagga tatgataatt
     5041 atggaggagg aaattacgga tctggaaatt acaatgattt cggaaactac aaccagcagc
     5101 catctaatta tggaccaatg aagtctggaa atttcggagg atctcgcaat atgggtggac
     5161 catatggagg aggaaattat ggtccaggag gatctggagg atctggagga tatggaggac
     5221 gttctcgtta ctaaattaat tccttaagcc cctctaagtg tccaacgtgt ctgagcttcc
     5281 cgtgctctct cctctgtatc gatcttctca accatttttg ctctgctatc tcatcaaaat
     5341 cttctgtact ttttcttgtt gcctttctcc agaaagtttc gtttattatt caatttttta
     5401 ccctgtcttt tttatataat ttatttctaa attcgcacag ttgacatcca ttcaactaga
     5461 gaatgaatgc agagtttgac accaaattgg cttaattttc aagctgtttt taaccaaaat
     5521 aataaaaaag tctcaacttt acaaaaatgt ttatctggat ctggacgaca gcgagctgag
     5581 aaattgatat attttcagag taaaaactaa aaatgtctgg aatttttgga gtcattgtct
     5641 cagggaggac gccgatcgaa gtggtgccgg tctccgatac ggagttcacc tgcgaaattg
     5701 taaacgctga cgcgataaac cacgtggtgg tcttccttac cggagctcaa ccgtttccgg
     5761 acggaattgg tggctctggt gatcattttc agaattttcc gcccaaaaaa acccaattta
     5821 tttcagttta catcagatgg cccacccaag acggcggaaa ttggcattat ttgggattta
     5881 tttgcaacga gaagcccagc gcgatttata aagttgctca ggtgtgcatt tttgatggaa
     5941 aaatgaaaga aaaccaccga aatttcgatg tttctctgtc gatttttttt atttaaaaac
     6001 ttagaaatac tctcaatttc gcatttttca ccaaaaaaaa gaatctcaac tggtttttca
     6061 catagcttaa tttttgaaga aaaatctcaa caaaacccag ttttaagttt caaattctca
     6121 aaaattgatc ttttttttcc cttaaaaatc cagtttttag ctccaaattg tcataatttt
     6181 ccagaaaaat ttagttttcg agctaacaac attaaaattt cacagaaaat ctgaatttca
     6241 gccttagaga tccaaatttt tcggctgaaa atcacaattt ttccattttt cagctccaca
     6301 aatccgacgc ttctcactcg ttttcccaat ttgacaatca aatgcaacta tattccagtg
     6361 gatcagctca aattggaatt aatgctgaaa gtttatctga tatcactgga cggcaggcag
     6421 ctgatgggac acaggctagc cagcaggtga aggatccccc gggctgcagg aattcgatat
     6481 caagcttatc gataccgtcg acctcgaggg ggggcccggt acccagcttt tgttcccttt
     6541 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6601 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6661 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6721 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6781 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6841 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6901 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6961 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7021 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7081 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7141 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7201 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7261 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7321 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7381 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7441 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7501 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7561 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7621 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7681 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7741 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7801 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7861 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7921 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7981 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8041 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8101 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8161 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8221 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8281 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8341 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8401 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8461 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8521 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8581 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8641 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8701 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     8761 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     8821 ttataaatca aaagaataga ccgagatagg gttgagtg
//
