LOCUS       Exported                5277 bp DNA     circular SYN 23-OCT-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5277)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5277
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..33
                     /label=TP59
     misc_feature    7..12
                     /label=NheI
     misc_feature    8..1160
                     /label=DONOR LEFT ARM
     CDS             join(198..494,1128..1314)
                     /codon_start=2
                     /label=Isorform B coding region
                     /translation="FLNNRSKRNSLSANAVATEQKATKVLGLVFFTFVLCWSPFFILNI
                     IFAACPECQVPEHVVNTCLWLGYVSSTINPIIYTIFNRTFRAAFIRLLKCNCKRSGRPL
                     RYRSVTEARGALSLCAPSALPLAISFQGAPLMTPSTANATPMSEFRASCRTVTDDEC"
     primer_bind     1111..1132
                     /label=P118
     misc_feature    1144..1163
                     /label=gRNA2-1 for BFH
     primer_bind     complement(1154..1173)
                     /label=MP17
     primer_bind     1154..1173
                     /label=MP18
     misc_feature    1161..1314
                     /label=INSERT UPSTREAM
     misc_feature    1164..1166
                     /label=PAM, AGG to AGC, G881A
     primer_bind     complement(1293..1322)
                     /label=TP60
     primer_bind     1305..1333
                     /label=TP61
                     /label=TP36
     misc_feature    1315..1845
                     /label=insert fragment
     CDS             1315..1359
                     /codon_start=1
                     /label=C-linker
                     /translation="GDGGSGGDGGSGGGS"
     primer_bind     complement(1317..1335)
                     /label=P32
     CDS             1360..1407
                     /codon_start=1
                     /label=GFP11
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1408..1440
                     /codon_start=1
                     /label=linker2
                     /translation="SGGDGGSGGGS"
     CDS             1441..1488
                     /codon_start=1
                     /label=GFP11
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1504..1551
                     /codon_start=1
                     /label=GFP11-COPY3
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1567..1614
                     /codon_start=1
                     /label=GFP11-COPY4
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1630..1677
                     /codon_start=1
                     /label=GFP11-COPY5
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1693..1740
                     /codon_start=1
                     /label=GFP11-COPY6
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1756..1803
                     /codon_start=1
                     /label=GFP11-COPY7
                     /translation="RDHMVLHEYVNAAGIT"
     primer_bind     1795..1823
                     /label=P84
     primer_bind     complement(1795..1823)
                     /label=P83
     primer_bind     complement(1795..1819)
                     /label=P75
     CDS             1807..1833
                     /codon_start=1
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1819..1856)
                     /label=TP63
                     /label=TP37
     primer_bind     1830..1866
                     /label=TP62
     misc_feature    1846..1928
                     /label=INSERT DOWNSTREAM
     primer_bind     complement(1906..1936)
                     /label=MP19
     primer_bind     1906..1936
                     /label=MP20
     misc_feature    1923..1925
                     /label=PAM, TGG to TGA
     misc_feature    complement(1926..1945)
                     /label=gRNA2-1 for isoform BFH
     misc_feature    1929..3046
                     /label=DONOR RIGHT ARM
     primer_bind     complement(1937..1962)
                     /label=P119
     primer_bind     complement(3026..3060)
                     /label=TP64
     primer_bind     complement(3042..3060)
                     /label=P117
     primer_bind     complement(3116..3132)
                     /label=M13 rev
     primer_bind     complement(3129..3150)
                     /label=M13R
     rep_origin      complement(3304..3892)
                     /direction=LEFT
                     /label=ori
     protein_bind    complement(4009..4033)
                     /label=attB1
     CDS             complement(4133..4993)
                     /label=AmpR
     promoter        complement(4994..5098)
                     /label=AmpR promoter
     primer_bind     5212..5228
                     /label=M13 fwd
ORIGIN
        1 aattatgcta gctgagtaac gggtatctga cagtcgagga acttgactat aacgttctct
       61 cttgtttata taactatggg tttctgatag atgtgacttt ctcgcgctct cactagatga
      121 gaaaataatg aaacaataat catcatggct taactttttg gtactactaa tatcttttat
      181 ccacttttgc cttacagttt tctaaacaat cgcagtaaac gaaacagctt gtcggccaat
      241 gccgttgcta cggagcagaa ggccaccaag gtgctcggat tggtgttttt caccttcgtt
      301 ctgtgctggt cgcctttctt catcttgaac attatatttg ccgcatgccc cgagtgccaa
      361 gtgccggagc acgtggtgaa tacctgcctt tggcttggct atgtctcctc gacgatcaat
      421 cccatcatct acaccatatt caatcgcacg ttccgggcgg cctttatccg cctgctcaag
      481 tgcaactgca aacggtgagt agtagagcag tgagaagtca ctgcagccca gacgcggatc
      541 cagtgccaca gtaagaaatt gcctctccca accgacgaag gcacctcgaa ctaaaattcg
      601 tcgctgcagg ttacagtgac acagggatgc aatttattgc tctaagagtg cattctgctg
      661 ggaattaatg tgcaacgtga cgtatgattg atttttgggg ttttgattta ccagaggaat
      721 ccagacaaga gacgaactcg gggtggttgt ggcagcgaat gaataagcag cgataggaaa
      781 tcaaacgctc tcaaaatata ttaaaggaag aataagttcg attgtttgta tttaatatgc
      841 agaatgaatg ggaatctttg taatacgact tttatctatt ttaaagactt gctagaatcg
      901 ccacacattg taagacataa atttcgtaag cattaaaaac aaactgtgtc gaatgtcaaa
      961 tttatttaaa atgccttgtg ttcgaattaa aaacgtattg aaaagtatag gacatttacc
     1021 aagacgagag cacttttctg tcactgactt ctgccttaaa acccaatcga ctcgaaatgg
     1081 cttaaaatgg cacagccaat aacgtagtta ttgccatccc attgcagttc cggccgcccg
     1141 ctgcgctatc ggtctgtgac agaagcacgc ggcgctctca gcctgtgcgc cccttcggcc
     1201 ctgccgctgg ccatatcctt ccagggggcg ccacttatga cgccatctac ggctaatgcg
     1261 acgccgatga gcgagttccg cgccagctgc cgcacagtca cagacgacga gtgcggcgat
     1321 ggaggaagcg gaggcgacgg aggcagcgga ggaggcagcc gggaccacat ggtgctgcac
     1381 gagtacgtga acgccgccgg catcactagc ggaggcgacg gaggcagcgg aggaggcagc
     1441 cgggaccaca tggtgctgca cgagtacgtg aacgccgccg gcatcactag cggtagtggc
     1501 ggccgtgacc acatggtcct tcatgagtat gtaaatgctg ctgggattac aggtggctct
     1561 ggaggtagag atcatatggt tctccacgaa tacgttaacg ccgcaggcat cactggcggt
     1621 agtggaggac gcgaccatat ggtactacat gaatatgtca atgcagccgg aataaccgga
     1681 gggtccggag gccgggatca catggtgctg catgagtatg tgaacgcggc gggtataact
     1741 ggtgggtcgg gcggacgaga ccatatggtg cttcacgaat acgtaaacgc agctggcatt
     1801 actagttacc catacgatgt tccagattac gcttaactct agacctgaac cacgaggagc
     1861 accatccagc gtctgggatg catcaaattt atatcgacgt atcggcagaa acaaaacgga
     1921 gttcactgta attagcggct tttcctagat aacttcaggg agatcaatgc taaccttttc
     1981 aatcacaatc aatttgatga attttgtaga tgctatagag aaaataccta acttaagtct
     2041 catcgcatat cattagaaac aaaggaaggt tcaaaaatat agtgagagag tttgcggtac
     2101 aacgcggact gtttttcact ttctcatcaa cattctagac ttggtcattg cgttatttct
     2161 aagtcaaaca aaaaatcatt aacaataagt acgattagtg caaaataaaa taagaaaaag
     2221 aaggtaaaag tattagtttt tttacccttt agtttatagt gggagtaaga gtcgttcgtt
     2281 atatgcaaat ttataaacaa attgtaatct tttcttttgc aaataagcaa aataaatgat
     2341 tccaataagc gccttttgct agcagctgga tctggaaaaa atatatttgt atacacttta
     2401 tcagtaaaaa cagccaggtc gatatagtcc gccttatacc ttttagttcg tagtagttta
     2461 aattggtagc atactcgtat aaatcggaac aggcagaatt ggatgcctac aacatacata
     2521 tatgcaactg ttataggaac gatcaaaaat caactttaaa gaaatgtatc tttatagtac
     2581 aaagctgtca tagtctgaga attactacct aattaatgga aagaaatctt agcttcgctg
     2641 ttttcataga taatatataa tttagtttcc cttttattga taaccagttt gtaagattcg
     2701 gctttctttt ttaattctac atagacatat attttacggc tagcctaatg aaaaaagttt
     2761 tgaaccaaca aaaaatccgc ttaaggatgt aatccttccg cacctcccac atacataagt
     2821 tccgatatca ccattgggtt gacaaattat tttttattca actaaaatat cctttgaaca
     2881 gagcgccggc ttttataaaa taaagtgaca cttgaatagg gaatattcat aatattaata
     2941 ctcgtaaata aatcaataca cgtattactc gtaataactc gtaaggaaga agggtatact
     3001 aaatactttg aaatatattt tacaggcaga aggaagcgtt tccgagcggc cgcaattata
     3061 ggctaggtgg aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat
     3121 agctgtttcc tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc
     3181 cgacaatctc caagacatta ggtggagttc agttcggcgt atggcatatg tcgctggaaa
     3241 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3481 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3721 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3901 tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca
     3961 gcgtaaatgg gtagggggct tcaaatcgtc ctcgtgatac caattcggag cctgcttttt
     4021 tgtacaaact tgttgataat ggcaattcaa ggatcttcac ctagatcctt ttaaattaaa
     4081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4261 caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5101 catttccccg aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc aaggaagtct
     5161 ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt tgtaaaacga
     5221 cggccagtca tgcataatcc gcacgcatct ggaaggaagt gccattccgc ctgacct
//
