LOCUS       Exported                4008 bp ds-DNA     circular SYN 21-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4008)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4008
                     /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"
     misc_feature    complement(481..498)
                     /label=SeqLa
     primer_bind     517..533
                     /label=LSLabs M13 FWD (-41)
     misc_feature    537..553
                     /label=M13 For -20
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     747..776
                     /label=oJDW 1529 H2A-mCHerry FWD + BamHI
     primer_bind     751..776
                     /label=oJDW 1017
     primer_bind     751..776
                     /label=oJDW 1017 (P2A_AvrII_H2A_FWD)
     CDS             757..1140
                     /codon_start=1
                     /label=H2A.F/Z
                     /translation="MAGGKAGKDSGKAKTKAVSRSQRAGLQFPVGRIHRHLKTRTTSHG
                     RVGATAAVYSAAILEYLTAEVLELAGNASKDLKVKRITPRHLQLAIRGDEELDSLIKAT
                     IAGGGVIPHIHKSLIGKKGQQKTA"
     primer_bind     757..776
                     /label=oJDW 1370
     CDS             1147..1854
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1292..1310)
                     /label=oJDW 253
     primer_bind     complement(1446..1467)
                     /label=oJDW 1778 mCherry Rev 3
     primer_bind     complement(1833..1854)
                     /label=oJDW 1371
     primer_bind     complement(1835..1869)
                     /label=oJDW 1018
     promoter        complement(1898..1915)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1920..2054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1998..2017
                     /label=SV40pA REVSeq
     protein_bind    complement(2117..2212)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2250..2268)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2269..2289)
                     /label=M13-rev
     primer_bind     complement(2269..2287)
                     /label=LSLabs M13 REV (-27)
     misc_feature    2303..2324
                     /label=SeqL B
     CDS             2402..3211
                     /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      3358..3946
                     /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtgggat cctaggatgg caggtggaaa agcaggtaaa
      781 gacagtggaa aagccaagac gaaagccgtc tccagatctc agagagctgg tctgcagttc
      841 cctgtaggtc gtatccacag gcacctgaag acacgaacca ccagccatgg ccgtgtcgga
      901 gctacagctg cggtttacag tgctgctatc ctggagtatc tcaccgctga ggtgctggag
      961 ttggctggaa acgcttctaa ggatctgaag gtgaagcgaa tcactcccag acatttacag
     1021 ctggccatcc gcggagacga ggagctggac tcgcttatca aggccaccat tgctggagga
     1081 ggagtcattc ctcacatcca caagtctctg attggtaaga agggccagca gaaaaccgca
     1141 ggggggatgg tgagcaaggg cgaggaggac aacatggcca tcatcaagga gttcatgcgc
     1201 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1261 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     1321 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1381 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1441 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1501 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1561 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1621 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1681 gacgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1741 aacgtcaaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1801 tacgagcgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaatga
     1861 tagctcgagc tcaagcttcg aattctgcag tctagaacta tagtgagtcg tattacgtag
     1921 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1981 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2041 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2101 tgtgggaggt tttttacaac tttattatac aaagttggca ttataaaaaa gcattgctta
     2161 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttggagctcc
     2221 atggtagcgt taacgcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc
     2281 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2341 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2401 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2461 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2521 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2581 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2641 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2701 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2761 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2821 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2881 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2941 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3001 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3061 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3121 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3181 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3241 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3301 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3361 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3421 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3481 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3541 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3601 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3661 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3721 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     3781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3961 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
