LOCUS       Exported                6370 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 6370)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 6, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pSA_0_mTagBFP2-T2A-sGFP1-10_synCoTC from 1 to 6369
            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..6370
                     /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..746
                     /locus_tag="B-actin exon 2 start (SA_1)"
                     /label=B-actin exon 2 start (SA_1)
     variation       744..788
                     /locus_tag="flexible linker (GGGGS)x3 zf-opt"
                     /label=flexible linker (GGGGS)x3 zf-opt
     variation       744..758
                     /locus_tag="(GGGGS)linker"
                     /label=(GGGGS)linker
     variation       759..773
                     /locus_tag="(GGGGS)linker(1)"
                     /label=(GGGGS)linker
                     /label=(GGGGS)linker(1)
     variation       774..788
                     /locus_tag="(GGGGS)linker(2)"
                     /label=(GGGGS)linker
                     /label=(GGGGS)linker(2)
     misc_feature    789..1487
                     /locus_tag="zf-mTagBFP2 (no TAA)"
                     /label=zf-mTagBFP2 (no TAA)
     variation       1488..1502
                     /locus_tag="(GGGGS)linker(3)"
                     /label=(GGGGS)linker
                     /label=(GGGGS)linker(3)
     misc_feature    1503..1544
                     /locus_tag="V5-tag zf-optim (GKPIPNPLLGLDST)"
                     /label=V5-tag zf-optim (GKPIPNPLLGLDST)
     protein_bind    1545..1598
                     /locus_tag="zf_opt-T2A"
                     /label=zf_opt-T2A
     misc_binding    1599..2243
                     /locus_tag="zf-sGFP1-10"
                     /label=zf-sGFP1-10
     misc_feature    2250..4124
                     /locus_tag="synCoTC"
                     /label=synCoTC
     misc_feature    2250..2334
                     /locus_tag="afp 3'UTR (truncated at polyA)"
                     /label=afp 3'UTR (truncated at polyA)
     terminator      2815..4124
                     /locus_tag="CoTC terminator element"
                     /label=CoTC terminator element
     primer_bind     complement(4145..4165)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     primer_bind     complement(4162..4184)
                     /locus_tag="m13/pUC Reverse"
                     /label=m13/pUC Reverse
     misc_binding    complement(4171..4193)
                     /locus_tag="LacO"
                     /label=LacO
     rep_origin      complement(4533..5215)
                     /direction=LEFT
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     misc_feature    complement(5310..6170)
                     /locus_tag="AmpR gene"
                     /label=AmpR gene
     CDS             complement(5313..5972)
                     /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 aggggaggag gaggatctgg aggaggagga tctggaggag
      781 gaggatctat gtccgaactt atcaaagaaa atatgcatat gaagctgtac atggagggaa
      841 ctgtggacaa ccaccacttc aagtgcacct ctgagggaga gggaaagcct tatgagggaa
      901 cccagaccat gcgcatcaag gtggtggagg gaggacctct gccttttgct tttgacatcc
      961 tggccacctc tttcctgtat ggaagcaaga ccttcatcaa ccacacccag ggaatccctg
     1021 acttcttcaa gcagtctttc cctgagggat tcacctggga aagagtgacc acctacgagg
     1081 atggaggagt gctgaccgcc acccaggaca cctctctcca ggatggatgc ctgatctaca
     1141 acgtgaagat cagaggagtg aactttacta gcaacggacc tgtgatgcaa aagaagaccc
     1201 tgggatggga ggctttcact gagaccctgt acccagctga tggaggactg gagggaagaa
     1261 atgacatggc tctgaagctg gtgggaggat ctcacctgat cgccaatgcc aagaccacct
     1321 acagaagcaa gaagcctgcc aagaacctga agatgcctgg agtgtactat gtggactaca
     1381 gactggagcg catcaaggag gccaacaatg agacctatgt ggagcagcat gaggtggctg
     1441 tggccagata ttgtgacctc cctagcaagc tgggacacaa gctgaatgga ggaggaggat
     1501 ctggaaagcc aatcccaaac cctctgctgg gactggacag caccgaggga agaggatctc
     1561 tcctgacctg tggagatgtg gaggagaacc ctggcccaat gagcaaggga gaggagctgt
     1621 ttactggagt ggtgccaatc ctggtggagc tggatggaga cgtgaatgga cacaaattct
     1681 ctgtgagagg agagggagag ggagatgcca ccattggaaa gctgaccctg aagttcattt
     1741 gtaccaccgg aaagctgcct gtcccttggc caaccctggt gaccaccctg acctatggag
     1801 tgcagtgctt cagcagatac cctgaccaca tgaagagaca tgacttcttc aagtctgcca
     1861 tgcctgaggg atatgtgcag gagagaacca tctctttcaa ggatgatgga aagtacaaga
     1921 ccagagctgt ggtgaagttt gagggagaca ccctggtgaa cagaattgag ctgaaaggca
     1981 ctgacttcaa ggaggatgga aacatcctgg gacacaagct ggagtacaac ttcaactctc
     2041 acaatgtgta catcaccgct gataagcaga agaatggcat caaggccaac ttcaccgtga
     2101 gacacaatgt ggaggatgga tctgtgcagc tggctgatca ctatcagcag aacaccccaa
     2161 ttggagacgg acctgtgctg ctgcctgata atcattacct gagcacccag actgtgctga
     2221 gcaaggaccc aaatgagaag taaactagta ttaagtctca gccaccgtta actgaacatg
     2281 tcaaaacctg tggagactgt tgagatttga tgttctgaaa agataaagcc tataaataaa
     2341 acaatactca aatacaaaag gtatcttttt taaaaagtta gtttactgct tccttaggct
     2401 ctttctaaat cccacccacc tgaaggacat caattatgga tttatccttc atgccttgta
     2461 taaatacaga tgtgcatatg aatttttatg ggttttttaa aaatgtggtt cattctatat
     2521 acataacaat tactttcaac atacataagc ctacctgtat ttgtgcatta tattacatgg
     2581 tacagataaa tagtcatgtg ccccatgaac attttggtca aagatagatt acagatacaa
     2641 tggtggtccc ataagattat gatgccatat ttttactgta cctcttcttg tgcctagata
     2701 cacaaatact gacttgtgtt ataattgcct gcagtattca ataatatggt gtatagattt
     2761 gtagcctacg agctataacc tatactatat agccaaggag tgcaggaggg tatgccatct
     2821 agatttgtgt gagtacactg atgtttgaac aataaaatca cctaatgata ggtttctcaa
     2881 accatatccc agtccttaag caacacatga ctaatggatt caaccatttc ccctgttcgt
     2941 tcagattgct ttttcttttt gcccctacaa ttttgttgat aatactgttt tttggggctt
     3001 ttgtttttgt tttttttttt ttttttgaga caagagtctc actctgtcgt caggctggag
     3061 tgcaatggcg ctatctcagc tcactgcaac ctctgcctcc caggttcaag cgattctcct
     3121 gcctcagcct cctgagtagc tgagattaca ggcatgcgcc accgcgcccg gctaattttg
     3181 tatttttagt agagacgcag tttcaccatg ttggtcaggc tggtcttgaa cacccgacct
     3241 caggtgatcc acccgcctca gcttcccaaa gtgctgggat taaaggcgtg agccaccaca
     3301 cctggccaat ttactgtttt ataccatgct tttttgtgtg tattctaggt tccctaaagc
     3361 aaacatgcat tttaaatttt gatagaacca gatgacatac aaaagtgcct gttttctctg
     3421 cattattaaa actgaaggta atcagtcttt tatttctatg actccaactg gcaaaaaata
     3481 tttgccattt taactttttt tttttgagac ggagtcaaaa ggtgccagtt ttctctgcat
     3541 tattaaaact gaaggtaatc agtcttttat ttctatgact ccaactggca aaagatattt
     3601 gccacgttaa cttttttttt tttttttttg agacggaggc tcacactgtc acccaggctg
     3661 aagtgcagtg gcatgatctc agctcacttg caacctccgc ctcccaggtt caagtgattc
     3721 tcctgcctca gcctcccaag gagttgggat tacaggcgca tacccggctt attttttgta
     3781 tttttagtag aaacgggatt tcaccgtgtt agccaggatg gtctcaatct ccagacttcg
     3841 tgatcgccca cctcggcctc ccaaagtgct gggattacag gcgtgagcca ccgcgcccgg
     3901 ccctcagtag gcgattttta tatgtggtat aaagtagaat tcttacagat ggatacccgt
     3961 ttatgccagt gccatgactg aaaccctgct tgtccaactt gaaatatctt caatgcccat
     4021 ctattcttcc ttgatcttgt tttttcctgt gataatacaa tagtttttac cagttaatca
     4081 gttaagatga ggcttaccaa atatatggga gtactactga acacgatatc aagcttggcg
     4141 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4201 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4261 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4321 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4381 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4441 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4501 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4561 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4621 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4681 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4741 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4801 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4861 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4921 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4981 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5041 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5101 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5161 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5221 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5281 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5341 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5401 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5461 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5521 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5581 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5641 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5701 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5761 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5821 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5881 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5941 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6001 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6061 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6121 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6181 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6241 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6301 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6361 ccctttcgtc
//
