LOCUS       Exported                3376 bp ds-DNA     circular SYN 21-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3376)
  AUTHORS   Wythe Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3376
                     /organism="recombinant plasmid"
                     /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"
     misc_feature    complement(481..498)
                     /label=SeqLa
     misc_feature    537..553
                     /label=M13 For -20
     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"
     primer_bind     645..728
                     /label=oJDW (Lifeact mScarlet FWD)
     primer_bind     670..708
                     /label=JDW 466
     primer_bind     682..728
                     /label=Primer 1
     regulatory      682..691
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             688..738
                     /codon_start=1
                     /label=Lifeact
                     /translation="MGVADLIKKFESISKEE"
     primer_bind     688..728
                     /label=F-AvrII-mScarlet
     regulatory      754..763
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             760..1455
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(893..911)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1161..1180
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1358..1378
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1423..1485)
                     /label=R-HA-AscI-XhoI-mScarlet
     primer_bind     complement(1428..1455)
                     /label=Primer 2
     primer_bind     complement(1456..1522)
                     /label=oJDW XXX (Lifeact mScarlet HA REV)
     CDS             1456..1482
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(1501..1600)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1618..1636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1641..1657)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1671..1692
                     /label=SeqL B
     CDS             1770..2579
                     /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      2726..3314
                     /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 caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcaac cggtggatcc agccaccatg ggcgtggccg acctgatcaa gaagttcgag
      721 agcatcagca aggaggaggg ggatccaccg gtcgccacca tggtgagcaa gggcgaggca
      781 gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag
      841 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      901 aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg
      961 tacggctcca gggccttcac caagcacccc gccgacatcc ccgactacta taagcagtcc
     1021 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc
     1081 gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc
     1141 accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc
     1201 accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc
     1261 ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc
     1321 gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag
     1381 gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg
     1441 gacgagctgt acaagtatcc atatgacgtt ccagattacg cttaataaac gcgtactagt
     1501 gcccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1561 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     1621 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1681 atctctgatg ttacattgca caagataaaa taatatcatc atgaacaata aaactgtctg
     1741 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     1801 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1861 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     1921 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     1981 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     2041 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     2101 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2161 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     2221 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2281 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     2341 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2401 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2461 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2521 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2581 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2641 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     2701 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2821 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2881 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2941 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3001 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3061 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3121 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3181 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3241 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3301 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3361 cttttgctca catgtt
//
