LOCUS       Exported                6155 bp ds-DNA     circular SYN 06-MAR-2023
DEFINITION  pUC57.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6155)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 6, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pSA_1_mTagBFP2-T2A-sGFP11x7_synCoTC from 1 to 6154
            pSA_0_mTagBFP2-T2A-sGFP11x7_synCoTC from 1 to 6153
            pSA_0_mTagBFP2_synCoTC from 1 to 5673
            pSA_0_mTagBFP2 from 1 to 4252
            F
            nnncctaggaGGCTGCTGCGGTTCCAGAGGTGGGAAGTTCCTATTCcGAAGTTCCTATTCTCTAGAA 
            AGTATAGGAACTTC
            R:
            GCCcctaggccaGAATGTAGTGGCCGATCTTCGAAGTTCCTATACTATTTGAAGAATAGGAACTTCg 
            GAATAGGAACTTCATCTAGATGCATTCGCGAGGT
FEATURES             Location/Qualifiers
     source          1..6155
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(239..307)
                     /locus_tag="LacZ alpha"
                     /label=LacZ alpha
     primer_bind     364..386
                     /locus_tag="m13/pUC Forward"
                     /label=m13/pUC Forward
     primer_bind     378..395
                     /locus_tag="M13-fwd"
                     /label=M13-fwd
     misc_feature    432..479
                     /locus_tag="48bp_FRT3"
                     /label=48bp_FRT3
     misc_feature    480..499
                     /locus_tag="sgH (human ampka1 guide)"
                     /label=sgH (human ampka1 guide)
     misc_feature    510..529
                     /locus_tag="sgM site"
                     /label=sgM site
     misc_feature    533..580
                     /locus_tag="48bp_FRT"
                     /label=48bp_FRT
     intron          581..742
                     /locus_tag="B-actin intron 3'"
                     /label=B-actin intron 3'
     exon            743..747
                     /locus_tag="B-actin exon 2 start (SA_2)"
                     /label=B-actin exon 2 start (SA_2)
     variation       745..789
                     /locus_tag="flexible linker (GGGGS)x3 zf-opt"
                     /label=flexible linker (GGGGS)x3 zf-opt
     variation       745..759
                     /locus_tag="(GGGGS)linker"
                     /label=(GGGGS)linker
     variation       760..774
                     /locus_tag="(GGGGS)linker(1)"
                     /label=(GGGGS)linker
                     /label=(GGGGS)linker(1)
     variation       775..789
                     /locus_tag="(GGGGS)linker(2)"
                     /label=(GGGGS)linker
                     /label=(GGGGS)linker(2)
     misc_feature    790..1488
                     /locus_tag="zf-mTagBFP2 (no TAA)"
                     /label=zf-mTagBFP2 (no TAA)
     variation       1489..1503
                     /locus_tag="(GGGGS)linker(3)"
                     /label=(GGGGS)linker
                     /label=(GGGGS)linker(3)
     misc_feature    1504..1545
                     /locus_tag="V5-tag zf-optim (GKPIPNPLLGLDST)"
                     /label=V5-tag zf-optim (GKPIPNPLLGLDST)
     protein_bind    1546..1599
                     /locus_tag="zf_opt-T2A"
                     /label=zf_opt-T2A
     misc_feature    1600..2028
                     /locus_tag="zf-sGFP11x7 (5aa linker)"
                     /label=zf-sGFP11x7 (5aa linker)
     misc_binding    1600..1647
                     /locus_tag="zf-GFP11(RDHMVLHEYVNAAGIT)"
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)
     misc_binding    1663..1710
                     /locus_tag="zf-GFP11(RDHMVLHEYVNAAGIT)(1)"
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)(1)
     misc_binding    1726..1773
                     /locus_tag="zf-GFP11(RDHMVLHEYVNAAGIT)(2)"
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)(2)
     misc_binding    1789..1836
                     /locus_tag="zf-GFP11(RDHMVLHEYVNAAGIT)(3)"
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)(3)
     misc_binding    1852..1899
                     /locus_tag="zf-GFP11(RDHMVLHEYVNAAGIT)(4)"
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)(4)
     misc_binding    1915..1962
                     /locus_tag="zf-GFP11(RDHMVLHEYVNAAGIT)(5)"
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)(5)
     misc_binding    1978..2025
                     /locus_tag="zf-GFP11(RDHMVLHEYVNAAGIT)(6)"
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)
                     /label=zf-GFP11(RDHMVLHEYVNAAGIT)(6)
     misc_feature    2035..3909
                     /locus_tag="synCoTC"
                     /label=synCoTC
     misc_feature    2035..2119
                     /locus_tag="afp 3'UTR (truncated at polyA)"
                     /label=afp 3'UTR (truncated at polyA)
     terminator      2600..3909
                     /locus_tag="CoTC terminator element"
                     /label=CoTC terminator element
     primer_bind     complement(3930..3950)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     primer_bind     complement(3947..3969)
                     /locus_tag="m13/pUC Reverse"
                     /label=m13/pUC Reverse
     misc_binding    complement(3956..3978)
                     /locus_tag="LacO"
                     /label=LacO
     rep_origin      complement(4318..5000)
                     /direction=LEFT
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     misc_feature    complement(5095..5955)
                     /locus_tag="AmpR gene"
                     /label=AmpR gene
     CDS             complement(5098..5757)
                     /locus_tag="AmpR"
                     /label=AmpR
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tgaagttcct attccgaagt tcctattctt caaatagtat aggaacttcg
      481 aagatcggcc actacattct ggcctaggag gctgctgcgg ttccagaggt gggaagttcc
      541 tattccgaag ttcctattct ctagaaagta taggaacttc aataaaagtt aatttaagcg
      601 ttcattacaa taaattcaag tgaatctgtt ttaagctagc ctttaacgtt tttaaattcc
      661 ttcaaacaaa ttggatcctc tttgcagttc ctcatttctc ggtgagctgg tgctttaatc
      721 cctttacttt ttcttcctct agggggagga ggaggatctg gaggaggagg atctggagga
      781 ggaggatcta tgtccgaact tatcaaagaa aatatgcata tgaagctgta catggaggga
      841 actgtggaca accaccactt caagtgcacc tctgagggag agggaaagcc ttatgaggga
      901 acccagacca tgcgcatcaa ggtggtggag ggaggacctc tgccttttgc ttttgacatc
      961 ctggccacct ctttcctgta tggaagcaag accttcatca accacaccca gggaatccct
     1021 gacttcttca agcagtcttt ccctgaggga ttcacctggg aaagagtgac cacctacgag
     1081 gatggaggag tgctgaccgc cacccaggac acctctctcc aggatggatg cctgatctac
     1141 aacgtgaaga tcagaggagt gaactttact agcaacggac ctgtgatgca aaagaagacc
     1201 ctgggatggg aggctttcac tgagaccctg tacccagctg atggaggact ggagggaaga
     1261 aatgacatgg ctctgaagct ggtgggagga tctcacctga tcgccaatgc caagaccacc
     1321 tacagaagca agaagcctgc caagaacctg aagatgcctg gagtgtacta tgtggactac
     1381 agactggagc gcatcaagga ggccaacaat gagacctatg tggagcagca tgaggtggct
     1441 gtggccagat attgtgacct ccctagcaag ctgggacaca agctgaatgg aggaggagga
     1501 tctggaaagc caatcccaaa ccctctgctg ggactggaca gcaccgaggg aagaggatct
     1561 ctcctgacct gtggagatgt ggaggagaac cctggcccaa gagaccacat ggtcctgcat
     1621 gagtacgtga acgccgctgg aattactgga ggatctggag gaagagacca catggtcctg
     1681 catgagtacg tgaacgccgc tggaattact ggaggatctg gaggaagaga ccacatggtc
     1741 ctgcatgagt acgtgaacgc cgctggaatt actggaggat ctggaggaag agaccacatg
     1801 gtcctgcatg agtacgtgaa cgccgctgga attactggag gatctggagg aagagaccac
     1861 atggtcctgc atgagtacgt gaacgccgct ggaattactg gaggatctgg aggaagagac
     1921 cacatggtcc tgcatgagta cgtgaacgcc gctggaatta ctggaggatc tggaggaaga
     1981 gaccacatgg tcctgcatga gtacgtgaac gccgctggaa ttacttaaac tagtattaag
     2041 tctcagccac cgttaactga acatgtcaaa acctgtggag actgttgaga tttgatgttc
     2101 tgaaaagata aagcctataa ataaaacaat actcaaatac aaaaggtatc ttttttaaaa
     2161 agttagttta ctgcttcctt aggctctttc taaatcccac ccacctgaag gacatcaatt
     2221 atggatttat ccttcatgcc ttgtataaat acagatgtgc atatgaattt ttatgggttt
     2281 tttaaaaatg tggttcattc tatatacata acaattactt tcaacataca taagcctacc
     2341 tgtatttgtg cattatatta catggtacag ataaatagtc atgtgcccca tgaacatttt
     2401 ggtcaaagat agattacaga tacaatggtg gtcccataag attatgatgc catattttta
     2461 ctgtacctct tcttgtgcct agatacacaa atactgactt gtgttataat tgcctgcagt
     2521 attcaataat atggtgtata gatttgtagc ctacgagcta taacctatac tatatagcca
     2581 aggagtgcag gagggtatgc catctagatt tgtgtgagta cactgatgtt tgaacaataa
     2641 aatcacctaa tgataggttt ctcaaaccat atcccagtcc ttaagcaaca catgactaat
     2701 ggattcaacc atttcccctg ttcgttcaga ttgctttttc tttttgcccc tacaattttg
     2761 ttgataatac tgttttttgg ggcttttgtt tttgtttttt tttttttttt tgagacaaga
     2821 gtctcactct gtcgtcaggc tggagtgcaa tggcgctatc tcagctcact gcaacctctg
     2881 cctcccaggt tcaagcgatt ctcctgcctc agcctcctga gtagctgaga ttacaggcat
     2941 gcgccaccgc gcccggctaa ttttgtattt ttagtagaga cgcagtttca ccatgttggt
     3001 caggctggtc ttgaacaccc gacctcaggt gatccacccg cctcagcttc ccaaagtgct
     3061 gggattaaag gcgtgagcca ccacacctgg ccaatttact gttttatacc atgctttttt
     3121 gtgtgtattc taggttccct aaagcaaaca tgcattttaa attttgatag aaccagatga
     3181 catacaaaag tgcctgtttt ctctgcatta ttaaaactga aggtaatcag tcttttattt
     3241 ctatgactcc aactggcaaa aaatatttgc cattttaact tttttttttt gagacggagt
     3301 caaaaggtgc cagttttctc tgcattatta aaactgaagg taatcagtct tttatttcta
     3361 tgactccaac tggcaaaaga tatttgccac gttaactttt tttttttttt ttttgagacg
     3421 gaggctcaca ctgtcaccca ggctgaagtg cagtggcatg atctcagctc acttgcaacc
     3481 tccgcctccc aggttcaagt gattctcctg cctcagcctc ccaaggagtt gggattacag
     3541 gcgcataccc ggcttatttt ttgtattttt agtagaaacg ggatttcacc gtgttagcca
     3601 ggatggtctc aatctccaga cttcgtgatc gcccacctcg gcctcccaaa gtgctgggat
     3661 tacaggcgtg agccaccgcg cccggccctc agtaggcgat ttttatatgt ggtataaagt
     3721 agaattctta cagatggata cccgtttatg ccagtgccat gactgaaacc ctgcttgtcc
     3781 aacttgaaat atcttcaatg cccatctatt cttccttgat cttgtttttt cctgtgataa
     3841 tacaatagtt tttaccagtt aatcagttaa gatgaggctt accaaatata tgggagtact
     3901 actgaacacg atatcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3961 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4021 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4081 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4141 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4501 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4621 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4741 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4981 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5041 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5101 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5161 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5221 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5281 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5341 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5401 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5461 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5521 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5581 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5641 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5701 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5761 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5821 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5881 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5941 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6001 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6061 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6121 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
