pENTRY-FRL1-mScarlet

LOCUS       pENTRY-FRL1-mScarlet               8770 bp DNA     circular SYN 28-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8770)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    28..127
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     145..161
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    182..3536
                     /label=FRL1
     CDS             3537..4232
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(5487..5503)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5528..5627)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6410..6943)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(7132..7160)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      7817..8405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(8540..8567)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(8659..8745)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttaacgcta gcatggatct cgggccccaa ataatgattt tattttgact gatagtgacc
       61 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      121 gcaggctggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt
      181 catcccggta aagcagattt agggatggtg cttagcaatg agaggtttca gcttcttctt
      241 gtcagcaaga atcttgagga agttgaagag gaaagggaca tagttgtgtt tccttctcat
      301 gttctctttc ttccagttct tagatttctc ctcctccatt acaatctttt gtgaaacagt
      361 ctctatcccg atattgactt ccgaaagctc tctatcaaga gcctcgtaac tgcttttctc
      421 agcgtattta tcagcttgca aagcacctaa ctgctgcaaa accctctctc gctttctttg
      481 atactctttc agttcagcta catacatctc tttcctgttc ttaactacag ctaagagact
      541 aaaccgaatc tcattctgcg aataccggtc aatctgctct tgtaccacag gttggaccat
      601 tctgagccac tcgattcctt ctggctcacc cagacactgt ccaagactaa tgggtccttc
      661 tttaagacca tcgagctcat ataagatacc atcaacaggt aagtagctga tgtaatgata
      721 cacatcatca tcttcgtcta gatttttcgc agcagctaat tcttcatctt ccatgatgga
      781 agacgggtca ggcctggcaa atgtgttgtg agctgcacgt attgcctcat tgttgttgat
      841 ggctaaacct ttcagttcag gaggaaattc tttggcgaat tgtttcagtt ctgatagttc
      901 tgagccaata tcaatactcg aagagttcat gaggactgat aatatcgctt gggtcgcaca
      961 agcattgttg attatctgca agaaacacat agtttataaa aaaggtaatg attggaaaat
     1021 ccgtgagtca caaatctaat gtgagagtaa aagacctggc ttgcaaagaa gaaattcggg
     1081 tttggctctg tgatgacaac acgattctcc ttttcttctg gacgccactt gtaaagcaat
     1141 atcaatccat agacaggtct gcaaatatca gacaactcaa ggtaaatttg actttgtttt
     1201 tcttctgttt cgaacaatag aagatgaaag agtaacaagt aaactacctt atttcatcca
     1261 gagagttgaa gtccaaggaa tacaattcct caacctgcac acctttcact tgcatttgtt
     1321 gtataatctc agtgaaaata cctaaaaata tagaatacag aaaaccaaaa aatatgaaga
     1381 aataacaaac aagaaagcga cattggtctt actcacatat atggcacaat ggctatcgag
     1441 agacactaat ccacttagta taagacaaac accattgttg atatataatt tcaaaaagca
     1501 tcaatacttt tattttccaa tcctcacaca gtaagtatca cctcttgtag atgcagagaa
     1561 gactattttc aacatttgtt ggcttctaat ttgacaaaat ccaaacataa atcagattct
     1621 acaggaagcc aagactattt tccaaacatt tttagtgctc cactatcaaa acctcgttac
     1681 tgacatcatc tctttatata tccagaaaac aggaacccca aattgttaaa gagaatatat
     1741 accaggatca gattctacag gaagccaaga catggttcaa cgatgactcg cgccggtgaa
     1801 aatgggtctg aatttagaat tttcactctc aaattcgcgc ttatctatct cctcccatcg
     1861 atttcatcgt cttcacgcta gctgcagact cgaaagagag actatagact ggtgtggtct
     1921 cgattttgtc ttaatagagg tcgctctaaa ccgagtagaa acgaaattga attcatccaa
     1981 accatttcaa tttctataaa tcgaaccaaa tcaagtgaaa aacaattcgg ttctggtttt
     2041 attaaactta ctcgaaccga accgaaccga taacgttctt atatatcctt gccggagaga
     2101 gccaaagaaa tcttagagat caaacaatga cggcgagtga gactatcgcc accgcgataa
     2161 atcagattga tgagaagaag gaaaagctaa agaaagcttt cgacgatctt caagctcatc
     2221 gctctctcct ttcaccgtct ttctcacttt cctggtctga gatcgattct cacttttcct
     2281 ctctccaatc ctcccttgct agccgtttcc gcctcctcca ctctacatct cctttggaac
     2341 atgattcgta tagaatcgac gcttctgatg cagggaaatc gtcgtcttcg gaggaggtat
     2401 cggagcaacc cgttgttgag ccggagttga gagccttgtg tgagaagatt gacggaattg
     2461 gtttgattaa gtatttgatt aggatatggg atgatgaaac tcctctcaat caggaagttt
     2521 ctgcagcgat tcgttactct ccggatacgg cgtccatggt gcttgacgcg attgaaggtt
     2581 cgaactacac accgtcatca tcgggaagat cattcgatgt aaggagagtt ttcgttttgt
     2641 taatggaagt tttgattgag atcaatgcga atatcactgt tgatacgaga aatagagcta
     2701 agaaactggc ttaccattgg aaatctaagg ttggtgtcaa gcctttcgag gcgttagtgt
     2761 ttctgcattt ggtcgcagct tttgaattag gatcagaatt tgacacagag gagctttctg
     2821 attacgtttt catgatcgct aagtataagc aagcaacttt ggtttgtaac aagattggtg
     2881 tagataggaa aagagttggg aaactcatca agactctctt ggattctggt aaaccaattt
     2941 tagcagtcaa attcatgtat gagtgtggga tgactgatga atttgagcca atccctgttt
     3001 tgaaatctta cattaaggac tgtagagaag ctgctcttag agtttgcgtg gaagataact
     3061 actctctcaa gtcgcagaac gaagcctctg ataaagaagt cagtgccttg aaaccattga
     3121 tcaaaatcat caaagatcag aaccttgaat cagagtttac tcaggagaaa gttgaggaac
     3181 gtgtagaaga gctggagaaa aataaggcat tgaggaaacg caacacgact aatcctccca
     3241 agcaagagcc acagcaaaag gggaaaaagc ggactagaga ctgcaaaaat gggtcacaag
     3301 ttccagttcc gagtcagcag ctactttcta gaccagaagc tcttcttatg cctgagcata
     3361 gtcatcatgg cttacagttg aacccttatg gtcttatgac ctcagcattt tctggtgttg
     3421 ttgtgaaccc attaactggg ctctttggtt caggtgcaac tcctcaatct ctttactatg
     3481 ctcagcaaac tggatatgtg ttgccgcctc aataccaccc gccttattat tctcagatgg
     3541 tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac atggagggct
     3601 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3661 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc tgggacatcc
     3721 tgtcccctca gttcatgtac ggctccaggg ccttcatcaa gcaccccgcc gacatccccg
     3781 actactataa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3841 acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc ctgatctaca
     3901 aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag aagaagacaa
     3961 tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg aagggcgaca
     4021 ttaagatggc cctgcgcctg aaggacggcg gccgctacct ggcggacttc aagaccacct
     4081 acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca
     4141 tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc gagggccgcc
     4201 actccaccgg cggcatggac gagctgtaca agtagctagc tgcatctcga caataagatc
     4261 ttaatctcct caatgtaaaa tgttatcttt gtagaatatt tgccttgatg gaaagtaaaa
     4321 aaatcattct aactctatgt ctgaggaatc tcattctttg tttttttggt ttatatagaa
     4381 tagtagtgaa tttttctaca atattggatc atgtttccta cattatagat tcttcgaaga
     4441 attcgcctac ccgcccgttg gcgtgtcttc aaacctcatc accttctatt tctagaggta
     4501 ccagtagcat atgcgcgagg aacgaggctc tcgacgggca ccttaatagc atcctttctc
     4561 ataccaatat ctgcaaatag ctgaatagct ttgctgccga aaggactaag ttgcgcaagc
     4621 cacgcagctg cactcactga gctggcttga ccgccccttc ccaacacata gtcttttccg
     4681 accagcaagc ttgttttaca atatattgca aaatcttcac aattgttttc caaagcatgg
     4741 taatcaccaa acccattccg caataagagg tatttggcgc gggaaatgac ttcatcgcag
     4801 ggatccgaag gtgctatagt gcatgtgccg cgtcgaggtt tggcgataaa tgcagctgga
     4861 gagacactgt actcgaaaag atagatgttt cctccggcga gaaagcagtc aaggcaagaa
     4921 gagattacac cttcgagatt caattgctct tcacaattag aacaaggatt gtctccttta
     4981 tgattaggga tgacactaac gacgatcttg tccaagaagg tcccagttcc agtttctaga
     5041 ccacctccac aagtgaaatg aatgactttc tcatcgccta tatatattcc tgaggatgag
     5101 catcaaatat ttgattatct ttgatgcaat ccgaaacaaa aaaaaagtca cgttattgag
     5161 agatagagac tgaaccgtga tgagagtaga tataagcgtt acgccatgaa tagatgtgat
     5221 ctcctggttt aagtttatct ctggagattt ggtttgagat aaatcctatc ttctccagaa
     5281 agtgggaatc cattttgatg actatctctc ttcctagctt tgttatggtt gtgattttat
     5341 aaccaaaatt taaagtcagc agccatattg tttttttacg tctgtagctg aatgtttctt
     5401 ctttattggc tttagtaaaa ggagtagtgg tattaaaata agagaaaatt tgatttcaga
     5461 gtcttcccat ctattgcagc ccgggggatc cactagttct agagcggccg ccaccgcggt
     5521 ggagctcacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     5581 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca
     5641 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     5701 atgaacaata aaactgtctg cttacataaa cagtaataca gggggtgtta tgagccatat
     5761 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     5821 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     5881 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     5941 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     6001 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     6061 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     6121 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     6181 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     6241 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     6301 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     6361 tgacgagggg aaattaggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg
     6421 tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag
     6481 ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt
     6541 tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc
     6601 cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac ctgggcagaa
     6661 cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg
     6721 tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg
     6781 ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt
     6841 cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag
     6901 ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct gctccataac
     6961 atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt
     7021 acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
     7081 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga
     7141 accgaacagg cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac
     7201 ccggcaacct tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc
     7261 aaggtttcgg tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag
     7321 gtgctgtgca cggatctgcc ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc
     7381 ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag
     7441 catcgtttgt tcgcccagga ctctagctat agttctagtg gttggctact aataggttgt
     7501 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     7561 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     7621 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     7681 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     7741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gctagccagg
     8521 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     8581 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     8641 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     8701 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     8761 ctactctcgc
//