LOCUS       Exported                5350 bp DNA     circular SYN 30-MAR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5350)
  AUTHORS   Emma Aleksandrovic
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2022 from SnapGene 6.0.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5350
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     terminator      73..100
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      192..278
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    331..430
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     407..447
                     /label=OTII_BP_F
     regulatory      433..442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             439..1146
                     /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(1200..1241)
                     /label=OTII_BP_R
     protein_bind    complement(1220..1319)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1431..1450)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1431..1450)
                     /label=Seq_pENTR_Rev
     CDS             1442..2251
                     /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"
     primer_bind     complement(1509..1528)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2394..2982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2883..2902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagcc aggaagagtt
       61 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
      121 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
      181 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
      241 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      301 cgcgttaacg ctagcatgga tctcgggccc caaataatga ttttattttg actgatagtg
      361 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa
      421 aaagcaggct tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
      481 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      541 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      601 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
      661 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
      721 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
      781 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
      841 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
      901 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
      961 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     1021 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     1081 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1141 tacaagtcag cagagagcct gaagatatct caagctgtcc atgcagcaca tgcagaaatc
     1201 aatgaagcag gcagataaga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     1261 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1321 catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1381 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1441 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1501 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1561 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1621 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1681 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1741 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1801 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1861 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1921 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     1981 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2041 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2101 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2161 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2221 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2281 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2341 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2401 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2461 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2521 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2581 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2641 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2701 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2761 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2821 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2881 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2941 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3001 tttttacggt tcctggcctt ttgctggcct tttgctcact ccctgctttg tatgaagaac
     3061 gccgttcgct gtcgtcaggt tgtggtcgac gcctcttgct gtctctgcct tggtctgtgc
     3121 ggtgtcgatg tgagctggac cccaactgtg gactgtcccc ttcagcttct ctatgcctgc
     3181 ttgacttgtg accatgcctg ttctcacgtg caactctatc gcggtcttgc ctgtgtgaat
     3241 gtgactttct ctctcgacgc tcatagctat gacggtccct gtctcgtctg ctgctatgtg
     3301 acggacttga ttcgcgaagc ggcgatgagg acctcctgtc ttttcggcaa gactcttcgc
     3361 ccttcgagct ggttgcctgc atcctcgact cggacatgat gatgttgata tcgagcacgt
     3421 tgatgggatc gatcctagaa tatcaatcga gccaccgtca atttcgagcg ctaagtggag
     3481 ttgtccgagt cccgaatcac aagaattagg ctcgtgctct gtgagatctc tcgcgaaacc
     3541 cagatgaagg aaaaaaatcc ggaagatcgc ggaagaaggt ggggttcgca tggtctaaca
     3601 ttgtcgcatt cctcacagtt tcggctggaa acggcaggga caatcacgag aaatgcgcga
     3661 cgagtattca gttgtccaaa attatgtcag cttgaaagtt ggaaacggcg gaagaaaatt
     3721 cgtcaggagg gcgttgcggc tagggtgaag aacggagaca atttcggctt tgaaattggc
     3781 tcgactctga ttggatcatg acgcactgct ggtccacaac cgtgtcgaag gtgccgtctt
     3841 tactacaggt ccgctggaag caaaatggaa aaagccgctg gagcccgact acagagccgc
     3901 cgtgttttgg taatcagtcg gcaaataggt cagcacagcg cagcgtgaca ggttcttgca
     3961 atttacagca cagctcgtcc gtctacgact ttgcacacca caaagtgtgc ggggagcaaa
     4021 ggagccgatc ttggtcgcgc gcaaagccaa ggagtcttga acctgagagt gtgcgtgtct
     4081 tgtgacgctt gcccttctgt actttgctgt gacactacca ccacatctgt cttggctttt
     4141 tgttcataca tccacaccga ccatgtcgct attcaacgtc agcaacggtc ttcgtaccgc
     4201 tctccgacct tctgttgcca gctcttcgcg cgttgctgcc ttttccacaa ccgccgctgc
     4261 ccgtctcgcc acacccacct ctgacaacgt tggcagttcg ggcaagcctc agcacttgaa
     4321 gcagttcaag atctaccgat ggaaccctga caagccctcg gagaagcctc gtctgcagtc
     4381 gtacacactg gacctcaacc agaccggtcc aatggttctc gacgcgctca tcaaaatcaa
     4441 gaacgaaatt gaccctacgc tcaccttccg tcgctcgtgc cgtgagggta tctgcggttc
     4501 gtgcgctatg aatattgacg gtgtcaacac cctcgcctgc ctctgccgga tcgacaagca
     4561 gaatgacacc aagatctacc ccttgccgca catgtacatt gtcaaggacc tcgtgccaga
     4621 cttgacccag ttctacaagc agtaccgatc catcgagcct ttcctcaagt ccaacaacac
     4681 cccttctgag ggtgaacatc ttcagtcgcc cgaggagcgt cgtcgactcg acggtctgta
     4741 cgagtgcatt ctgtgcgcgt gctgctccac atcctgcccc tcttactggt ggaatcagga
     4801 cgagtacctt ggccccgccg tgctcatgca ggcgtaccga tggatggccg actcgcgtga
     4861 cgactttggt gaggagcgaa gacagaagct cgagaacacc ttttcgctct accgatgcct
     4921 taccatcatg aactgctcca ggacctgccc caagaacctc aaccctggta aggcaattgc
     4981 acagatcaag aaggacatgg ccgtcggcgc acccaaggct tccgagcgcc ctatcatggc
     5041 ttcgtcgtaa tcttgatata tcatatcgtt ctttcctcag cacttctttt gtcaatttca
     5101 aaagtatcta attgcattca actccgcttg tggtttgttg ttcagtgaga gtggaaacgc
     5161 tacgggcaag atgagggcag tgttctggcg acggaaaagt gtgcaagtgt ctggcctgcg
     5221 tcctcgctgg ttccagcagc cgatgcagga cgtgtaccta gcgatttctt cgacagccta
     5281 ttgtggcagc cgcgattcgc cacaatcgta cgaaagcgag acgagttgag cgaagatccc
     5341 cgggcgagtt
//
