LOCUS       Exported                3812 bp ds-DNA     circular SYN 15-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3812)
  TITLE     Shinoda H, et al. Acid-tolerant monomeric GFP from Olindias formosa.
            Cell Chem. Biol. 2018;25:330?338.
REFERENCE   2  (bases 1 to 3812)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3812
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      681..690
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             687..689
                     /codon_start=1
                     /translation="M"
     CDS             690..731
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             738..761
                     /codon_start=1
                     /label=Linker
                     /translation="GSGGSGAS"
     CDS             762..1442
                     /codon_start=1
                     /label=oxStayGold(c4)_v2
                     /translation="MVSTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAYQNTIIKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHIIQSETLEAHLPWHEP
                     SASAV"
     misc_feature    771..1412
                     /label=StayGold (aa 4-X)
                     /note="missing amino terminal MAS of MAST sequence"
     misc_feature    1280..1283
                     /label=Cys>Ile
                     /note="cys->Ile for stability"
     misc_feature    1383..1385
                     /label=Cys>Ile
                     /note="Cys->Ile for stability"
     misc_feature    1413..1442
                     /label=C-term of dfGFP
                     /note="C-terminal of acid stable GFP (aa 223?233)"
     misc_feature    1415
                     /label=A>T
                     /note="silent mutation to destroy NcoI site"
     CDS             1443..1501
                     /codon_start=1
                     /label=GGGGSx3
                     /translation="EFDGGGGSGGGGSGGGGSP"
     misc_feature    1502..1877
                     /label=Giantin tag
     CDS             1502..1877
                     /codon_start=1
                     /label=Giantin_v5
                     /translation="SFSEAQQQLCNTRQEVNELRKLLEEERDQRVAAENALSVAEEQIR
                     RLEHSEWDSSRTPIIGSCGTQEQALLIDLTSNSCRRTRSGVGWKRVLRSLCHSRTRVPL
                     LAAIYFLMIHVLLILCFTGHL"
     protein_bind    complement(1937..2036)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2077..2093)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2206..3015
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3162..3750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg atccactagt gccaccatgg gtaagcctat ccctaaccct ctcctcggtc
      721 tcgattctac gggtaccgga tcaggaggat caggagctag catggtgagc acccccttca
      781 agttccagct gaagggcacc atcaacggca agagcttcac cgtggaaggc gagggcgagg
      841 gcaatagcca cgagggcagc cacaaaggca agtacgtgtg caccagcggc aaactgccaa
      901 tgtcttgggc cgccctggga actagcttcg gctatggcat gaagtactac accaagtacc
      961 ccagcggcct gaagaactgg ttccacgagg tgatgcccga gggcttcacc tacgacagac
     1021 acatccagta caagggcgac ggcagcatcc acgccaagca ccagcacttc atgaagaacg
     1081 gcacctacca caacatcgtg gagttcaccg gccaggactt caaggagaac agccccgtgc
     1141 tgaccggcga catgaacgtg agcctgccca acgaggtgca gcacatcccc agagatgacg
     1201 gcgtggagtg cccagtgacc ctgctgtacc ctctgctgag cgacaagagc aagtgcgtgg
     1261 aggcctacca gaacaccatc atcaagcccc tgcacaatca gccagccccc gatgtgccat
     1321 accactggat cagaaagcag tacacccaga gcaaggacga caccgaggag agagaccaca
     1381 tcatccagag cgagaccctg gaggcccacc tgccttggca cgagccttct gcttctgccg
     1441 tggaattcga tggtggaggc ggttcaggcg gaggtggctc tggcggtggc ggatcgccat
     1501 caagcttttc tgaagctcag cagcagctat gcaacaccag acaggaagtg aatgaattaa
     1561 ggaagctgct ggaagaagaa cgagaccaaa gagtggctgc tgagaatgct ctctctgtgg
     1621 ccgaggagca gatcagacgg ttagagcaca gtgaatggga ctcttcccgg actcctatca
     1681 ttggctcctg tggcactcag gagcaggcac tgttaataga tcttacaagc aacagttgtc
     1741 gaaggacccg gagtggcgtt ggatggaagc gagtcctgcg ttcactctgt cattcacgga
     1801 cccgagtgcc acttctagca gccatctact ttctaatgat tcatgtcctg ctcattctgt
     1861 gttttacggg ccatctatga tatctataat gaggcgcgcc gtcgacgtta acacgcgtta
     1921 atgatacgta tctagaaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     1981 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     2041 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     2101 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     2161 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     2221 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2281 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     2341 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2401 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2461 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     2521 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2581 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2641 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2701 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2761 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2821 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2881 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2941 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     3001 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     3061 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     3121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3301 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3781 ctggcctttt gctggccttt tgctcacatg tt
//
