LOCUS       Exported                4244 bp ds-DNA     circular SYN 15-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   urn.local...j-7twiatv
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4244)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 17, 2021 from SnapGene Viewer 5.2.5
            https://www.snapgene.com
COMMENT     SerialCloner_Ends=0,0,,0,.
FEATURES             Location/Qualifiers
     source          1..4244
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..57
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    1..57
                     /label=lastNN
                     /note="/created_by=User"
     misc_feature    58..180
                     /label=C-term region_Mok2020
                     /note="/modified_by=Issei.N"
     misc_feature    58..177
                     /label=C-term region_wako compact
                     /note="/Transferred_From=pH7WG2DIPS_ORF79cTAL14.xdna,pH7WG2
                     DIPS_ORF79cTAL8.xdna, pK7WG2DIPSHSP_ORF125_1.xdna, 
                     pK7WG2DIPSHSP_ORF125_2.xdna, 【E1】pENTR_L1-L4_HD, 
                     【E1】pENTR_L1-L4_NG, 【E1】pENTR_L1-L4_NI, 【E1】pENTR_L1-L4_NN,
                     【完成】pK7WG2DIPSHSP_platinum psbA, 【完成】pK7WG2DIPSHSP_platinum
                     これにLeftRight亘シークエンスをはればよい．"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=[mitoTAL]RPS5A-Shaorf117-1"
                     /note="/X-Transferred_Similarity=100.00%"
     primer_bind     107..126
                     /label=TALR1r
     primer_bind     complement(107..126)
                     /label=TALR2
     primer_bind     complement(161..180)
                     /label=E1E3_Rv
     primer_bind     181..200
                     /label=G1397-N_Fw
     misc_feature    181..186
                     /label=2aa linker
                     /note="/created_by=Issei.N"
                     /note="/modified_by=Issei.N"
     misc_feature    182..771
                     /label=ATCCTATCAGCCGTTCCCAGCTGGTGAAGTCCGAGCTGGAGGAGAAG...
                     /note="Geneious type: Editing History Replacement"
                     /note="/Original_Bases=ATCCTATCAGCCGTTCCCAGCTGGTGAAGTCCGAGC
                     TGGAG 
                     GAGAAGAAATCCGAGTTGAGGCACAAGCTGAAGTACGTGCCCCACGAGTACATCGAGC 
                     TGATCGAGATCGCCCGGAACAGCACCCAGGACCGTATCCTGGAGATGAAGGTGATGGA 
                     GTTCTTCATGAAGGTGTACGGCTACAGGGGCAAGCACCTGGGCGGCTCCAGGAAGCCC 
                     GACGGCGCCATCTACACCGTGGGCTCCCCCATCGACTACGGCGTGATCGTGGACACCA 
                     AGGCCTACTCCGGCGGCTACAACCTGCCCATCGGCCAGGCCGACGAAATGCAGAGGTA 
                     CGTGGAGGAGAACCAGACCAGGAACAAGCACATCAACCCCAACGAGTGGTGGAAGGTG 
                     TACCCCTCCAGCGTGACCGAGTTCAAGTTCCTGTTCGTGTCCGGCCACTTCAAGGGCA 
                     ACTACAAGGCCCAGCTGACCAGGCTGAACCACATCACCAACTGCAACGGCGCCGTGCT 
                     GTCCGTGGAGGAGCTCCTGATCGGCGGCGAGATGATCAAGGCCGGCACCCTGACCCTG 
                     GAGGAGGTGAGGAGGAAGTTCAACAACGGCGAGATCAACTTCGCGGCCGAC"
     misc_feature    187..510
                     /label=G1397-DddtoxA-N
                     /note="/created_by=Issei.N"
                     /note="/modified_by=Issei.N"
     misc_feature    511..522
                     /label=4aa linker
                     /note="/created_by=Issei.N"
     misc_feature    523..771
                     /label=UGI
                     /note="Derived using Geneious Prime 2020.1.2 'Annotatefrom 
                     Database' based on nucleotide similarity"
                     /note="/Alternative_Match=79.52%; Adjusted-Interval-0:0/0; 
                     UGI(<a href=""urn:local:.:5k-c662uqm"">2aa linker + 
                     G1333-DddtoxA-C + 4aa linker + 1*UGI     </a>)"
                     /note="/Primary_Match=79.92%; Adjusted-Interval-0:0/0; UGI 
                     (<a href=""urn:local:.:5s-c662uqm"">2aa linker + 
                     G1397-DddtoxA-C + 4aa linker + 1*UGI </a>)"
                     /note="/Transferred_From=2aa linker + G1397-DddtoxA-C + 
                     4aalinker + 1*UGI "
                     /note="/Transferred_Similarity=79.92%"
                     /note="/X-Alternative_Match=79.52%; 
                     Adjusted-Interval-0:0/0; UGI(<a 
                     href=""""urn:local:.:5k-c662uqm"""">2aa linker + 
                     G1333-DddtoxA-C + 4aa linker + 1*UGI     </a>)"
                     /note="/X-Primary_Match=83.13%; Adjusted-Interval-0:0/0; 
                     UGI (<a href=""""urn:local:.:5t-c662uqm"""">2aa linker + 
                     G1333-DddtoxA-N + 4aa linker + 1*UGI     </a>)"
                     /note="/X-Transferred_From=2aa linker + G1333-DddtoxA-N + 
                     4aalinker + 1*UGI     "
                     /note="/X-Transferred_Similarity=83.13%"
                     /note="/created_by=Issei.N"
     primer_bind     complement(751..771)
                     /label=UGI_Rv_G1397-N
     primer_bind     772..791
                     /label=E1E3_Fw
     misc_recomb     796..820
                     /label=attB4
                     /note="/Transferred_From=【完成】pK7WG2DIPSHSP_platinum psbA, 
                     【完成】pK7WG2DIPSHSP_platinum これにLeftRight亘シークエンスをはればよい．"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=[mitoTAL]RPS5A-Shaorf117-1"
                     /note="/X-Transferred_Similarity=100.00%"
     misc_recomb     complement(797..895)
                     /label=attL4
     misc_recomb     797..820
                     /label=attB4
     misc_feature    complement(800..896)
                     /label=nonstandard type: attL4
     misc_feature    complement(800..820)
                     /label=nonstandard type: attB4
                     /note="/Transferred_From=【完成】pK7WG2DIPSHSP_platinum psbA, 
                     【完成】pK7WG2DIPSHSP_platinum これにLeftRight亘シークエンスをはればよい．"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=[mitoTAL]RPS5A-Shaorf117-1"
                     /note="/X-Transferred_Similarity=100.00%"
     misc_recomb     complement(804..820)
                     /label=attR4
                     /note="/Transferred_From=【完成】pK7WG2DIPSHSP_platinum psbA, 
                     【完成】pK7WG2DIPSHSP_platinum これにLeftRight亘シークエンスをはればよい．"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=[mitoTAL]RPS5A-Shaorf117-1"
                     /note="/X-Transferred_Similarity=100.00%"
     misc_recomb     complement(810..820)
                     /label=attR3
                     /note="/Transferred_From=【完成】pK7WG2DIPSHSP_platinum psbA, 
                     【完成】pK7WG2DIPSHSP_platinum これにLeftRight亘シークエンスをはればよい．"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=[mitoTAL]RPS5A-Shaorf117-1"
                     /note="/X-Transferred_Similarity=100.00%"
     misc_recomb     810..820
                     /label=attB3
                     /note="/Transferred_From=【完成】pK7WG2DIPSHSP_platinum psbA, 
                     【完成】pK7WG2DIPSHSP_platinum これにLeftRight亘シークエンスをはればよい．"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=[mitoTAL]RPS5A-Shaorf117-1"
                     /note="/X-Transferred_Similarity=100.00%"
     primer_bind     complement(920..939)
                     /label=T7
     primer_bind     complement(940..960)
                     /label=M13-rev
     CDS             1073..1879
                     /label=KanR
     rep_origin      2007..2635
                     /direction=RIGHT
                     /label=ColE1 origin
     misc_RNA        complement(2068..2175)
                     /label=RNAI transcript
                     /note="RNAI transcript"
                     /note="/Transferred_From=pK7WG2CTP_RPS5Apro 
                     ver2,pK7WG2DIPS_RPS5Apro ver2, pK7WG2_pRPS5A_CTP, 
                     sequence後：pK7WG2_pRPS5A_CTP, 【完成】pK7WG2DIPSHSP_platinum 
                     psbA"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=DP1 GFP vector 
                     confirmed;pK7WG2CTP_RPS5Apro ver2; pK7WG2DIPS_RPS5Apro 
                     ver2; pK7WG2_pRPS5A_CTP; sequence後：pK7WG2_pRPS5A_CTP; 
                     【DEST】pK7WG2DIPS_RPS5Apro - コピー.xdna"
                     /note="/X-Transferred_Similarity=100.00%"
     misc_feature    2717..2736
                     /label=VR
                     /note="Geneious type: primer_bind_reverse"
                     /note="/%GC=50.0"
                     /note="/Hairpin_Tm=None"
                     /note="/Mismatches=0"
                     /note="/Pair_Dimer_Tm=None"
                     /note="/Self_Dimer_Tm=None"
                     /note="/Sequence=ATTACCGCCTTTGAGTGAGC"
                     /note="/Tm=58.3"
                     /note="/Transferred_From=pK7WG2_pRPS5A_CTP,【完成】pK7WG2DIPSHS
                     P_platinum psbA"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=DP1 GFP vector 
                     confirmed;pK7WG2_pRPS5A_CTP; pMDCDIPS20181026作製途中ほぼ完成？; 
                     【DEST】pK7WG2DIPS_RPS5Apro - コピー.xdna"
                     /note="/X-Transferred_Similarity=100.00%"
                     /note="/created_by=primer3"
                     /note="/modified_by=User"
     primer_bind     2717..2736
                     /label=VR
     primer_bind     2771..2788
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="/Transferred_From=pK7WG2_pRPS5A_CTP"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=【DEST】pK7WG2DIPS_RPS5Apro - 
                     コピー.xdna"
                     /note="/X-Transferred_Similarity=100.00%"
     primer_bind     2771..2788
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="/Transferred_From=pK7WG2_pRPS5A_CTP"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=Exported"
                     /note="/X-Transferred_Similarity=100.00%"
     primer_bind     2771..2788
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="/Transferred_From=【完成】pK7WG2DIPSHSP_platinum psbA"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/X-Transferred_From=pK7WG2_pRPS5A_CTP"
                     /note="/X-Transferred_Similarity=100.00%"
     terminator      complement(2947..2974)
                     /label=rrnB T2
     terminator      complement(3106..3149)
                     /label=rrnB T1
     primer_bind     3188..3207
                     /label=E1E3Ntercheck_Fw
     primer_bind     3215..3232
                     /label=M13-fwd
     misc_feature    3248..3344
                     /label=nonstandard type: attL1
     misc_recomb     3249..3344
                     /label=AttL1
     CDS             3350..3376
                     /label=HA tag
     misc_feature    3383..3790
                     /label=N-term region_wako; N-term region_wako compact
                     /note="/Transferred_From=pK7WG2DIPSHSP_ORF125_1.xdna,pK7WG2
                     DIPSHSP_ORF125_2.xdna, pH7WG2DIPS_ORF79cTAL14.xdna, 
                     pH7WG2DIPS_ORF79cTAL8.xdna"
                     /note="/Transferred_Similarity=99.75% -> 99.26%"
     primer_bind     complement(3603..3622)
                     /label=TALF1r
     primer_bind     3603..3622
                     /label=TALF1
     CDS             complement(3818..3886)
                     /label=LacZ alpha
     primer_bind     3957..3974
                     /label=M13-fwd
     primer_bind     3984..4003
                     /label=T7
     primer_bind     complement(4048..4067)
                     /label=T3
     primer_bind     complement(4085..4105)
                     /label=M13-rev
     primer_bind     4102..4123
                     /label=E1E3-check_Fw
     misc_binding    complement(4111..4133)
                     /label=LacO
     promoter        complement(4138..4167)
                     /label=lac
     misc_feature    join(4170..4244,1..774)
                     /label=ORF 4 (frame 3)
                     /label=nonstandard type: ORF
                     /note="/Genetic_code=Standard"
                     /note="/Reading_frame=3"
                     /note="/created_by=ORF Finder"
ORIGIN
        1 acccctgagc aggtagtggc tattgcatcc aacaacgggg gcagacccgc actggagtca
       61 attgttgccc agttatctcg ccctgatccg gcgttggccg cgttgaccaa cgaccacctc
      121 gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg cagtgaaaaa gggattgggg
      181 gggtctggat cgtatgcttt aggaccgtat cagatctcag ctccacaatt gcctgcatat
      241 aacggacaaa ctgttgggac cttttactac gttaacgatg ctggtggatt ggagtccaaa
      301 gtgttctctt ctggtggccc aactccatat cccaattatg cgaatgcagg ccatgttgaa
      361 ggtcaatcag ccctattcat gagagataac ggaataagtg aaggactggt gtttcacaac
      421 aatccagaag gtacttgtgg attttgcgta aacatgactg agacacttct cccagaaaat
      481 gccaagatga cagttgtacc tcctgaaggt tctggtggat cgacaaacct ttcagacatt
      541 atcgagaaag agacaggcaa acagctagtg attcaagagt ccattctcat gcttcccgaa
      601 gaagttgagg aagtcattgg gaataagccg gaaagtgaca tactcgttca tacggcttac
      661 gatgagagca cggatgagaa tgtcatgttg cttaccagtg atgcacctga atacaaacct
      721 tgggctttag tcatccagga cagcaatggt gagaacaaga tcaagatgct gtgataactc
      781 gagcgatcct ctagacaccc aacttttcta tacaaagttg gcattataag aaagcattgc
      841 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
      901 ccagctgcag ggcggccgcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
      961 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1021 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     1081 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     1141 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     1201 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1261 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     1321 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     1381 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     1441 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     1501 cgtatttcgc cttgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     1561 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     1621 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     1681 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     1741 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     1801 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     1861 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     1921 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     1981 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2041 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2101 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2161 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2221 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2281 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2341 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2401 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2461 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2521 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2581 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2641 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2701 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2761 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     2821 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     2881 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     2941 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     3001 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     3061 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     3121 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     3181 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
     3241 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     3301 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctttt acccatacga
     3361 tgttccagat tacgctgata tcgccgatct acgcacgctc ggctacagcc agcagcaaca
     3421 ggagaagatc aaaccgaagg ttcgttcgac agtggcgcag caccacgagg cactggtcgg
     3481 ccacgggttt acacacgcgc acatcgttgc gttaagccaa cacccggcag cgttagggac
     3541 cgtcgctgtc aagtatcagg acatgatcgc agcgttgcca gaggcgacac acgaagcgat
     3601 cgttggcgtc ggcaaacagt ggtccggcgc acgcgctctg gaggccttgc tcacggtggc
     3661 gggagagttg agaggtccac cgttacagtt ggacacaggc caacttctca agattgcaaa
     3721 acgtggcggc gtgaccgcag tggaggcagt gcatgcatgg cgcaatgcac tgacgggtgc
     3781 cccgctcaag gagacgggcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     3841 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     3901 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     3961 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc
     4021 cccccctcga ggccctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     4081 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4141 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4201 ttaattgcgt tgcgctcact gcccgctttc caccggtcgt ctcc
//
