LOCUS       Exported                3827 bp ds-DNA     circular SYN 10-MAY-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3827)
  AUTHORS   Wythe Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 10, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3827
                     /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"
     misc_feature    681..686
                     /label=Kozak
     CDS             687..1063
                     /codon_start=1
                     /label=H. sapiens (human) Histone cluster 1, H2bb, H2B
                     /translation="MPEPSKSAPAPKKGSKKAITKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSS"
     misc_feature    878
                     /label=should be a T(silent mutation)?
     CDS             1065..1079
                     /codon_start=1
                     /product="Gly Gly Ala Gly Gly Flexible Linker"
                     /label=G2AG2 Linker
                     /label=3X Flag GlyGlyAlaGlyGly
                     /note="Sabourin M et al., A flexible protein linker 
                     improves the function of epitope-tagged proteins in 
                     Saccharomyces cerevisiae (2007). Yeast."
                     /translation="GGAGG"
     CDS             1080..1121
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1122..1133
                     /codon_start=1
                     /label=MCS
                     /translation="GSTG"
     CDS             1134..1192
                     /codon_start=1
                     /label=GGGGS(x3)
                     /translation="EFDGGGGSGGGGSGGGGSP"
     CDS             1209..1895
                     /codon_start=1
                     /label=mScarlet3-H
                     /translation="GGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG
                     GGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG
                     GGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG
                     GGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG
                     GGGGGGG"
     regulatory      1209..1212
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(1952..2051)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2092..2108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2221..3030
                     /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      3177..3765
                     /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 agcaggctac tagtgctagc gccaccatgc ctgaaccctc taagtctgct ccagccccta
      721 aaaagggttc taagaaggct atcactaagg cgcagaagaa ggatggtaag aagcgtaagc
      781 gcagccgcaa ggagagctat tctatctatg tgtacaaggt tctgaagcag gtccaccccg
      841 acaccggcat ctcatccaag gccatgggga tcatgaactc cttcgtcaac gacatcttcg
      901 agcgcatcgc gggcgaggct tctcgcctgg ctcactacaa taagcgctcg accatcacct
      961 ccagggagat tcagacggct gtgcgcctgc tgctgcctgg ggagctggct aagcatgctg
     1021 tgtccgaggg cactaaggca gttaccaagt acactagctc taagggtggt gctggtggtg
     1081 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gggatccacc ggtgaattcg
     1141 atggtggagg cggttcaggc ggaggtggct ctggcggtgg cggatcgcca tcaggtacca
     1201 tggtgagcat ggatagcacc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca
     1261 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1321 acgagggcac ccagaccgcc aagctgaggg tgaccaaggg tggccccctg cccttctcct
     1381 gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcacgaag caccccgccg
     1441 acatccccga ctactggaag cagtccttcc ccgagggctt caagtgggag cgcgtgatga
     1501 acttcgagga cggcggcgcc gtgtccgtgg cccaggacac ctccctggag gacggcaccc
     1561 tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga
     1621 agaagacaat gggctgggaa gcatccaccg agcggttgta ccccgaggac gtcgtgctga
     1681 agggcgacat taagcatgcc ctgcgcctga aggacggcgg ccggtacctg gcggacttca
     1741 agaccaccta cagggccaag aagcccgtgc agatgcccgg cgccttcaac atcgaccgca
     1801 agttggacat cacatcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg
     1861 tggcccgcca ctccaccggc ggctccggtg gctcctaact ataatgaggc gcgccgtcga
     1921 cgttaacacg cgttaatgat acgtatctag aacccagctt tcttgtacaa agttggcatt
     1981 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     2041 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     2101 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     2161 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2221 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2281 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2341 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2401 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2461 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2521 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2581 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2641 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2701 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2761 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2821 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2881 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2941 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3001 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     3061 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     3121 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3301 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
