pENTRY-Col-0 fri-GFP

LOCUS       pENTRY-Col-0 fri-GFP                8660 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 8660)
  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..8660
                     /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..2510
                     /label=Col-0 fri
     CDS             2520..3236
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5377..5393)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5418..5517)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6300..6833)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(7022..7050)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      7707..8295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(8430..8457)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(8549..8635)
                     /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 cttgcccaat cactcgtggt aggatttttt tataagaaag ttacgaaaat gccctctcgt
      241 gcttagagag attcaattcg tgcacatata atatgccgtg aatataccac caactctctt
      301 actcgtactc gcatgtggtg cgtgcaatat acacgtggtt cgaatcaacg gctacgtttt
      361 actgataggt ggctcgttta gttttgcact gcctttgggt gcgcgtgacc aagtcatttt
      421 ccaaatgtgt tttaatattt gtgaactatt attaataaaa aagcgagcga cgttgttcgt
      481 tgatgaggga gcgaccgtac gcttggcgac cttctctgat tgcgtagagt cgttttcctt
      541 attgggctta cctagtaccg tcagcccaca atgttttcat ttctttccca cctaaaacta
      601 aaagtactca caagtcacaa cttaaaccaa gtacacaagg attttatcat gggattatcg
      661 tgtttgaaga ctaaaaagag cacaccatca cccccattag tgcaggtaga gtaagacagt
      721 aacttttggg ttcatattac cgagcaagaa ccgttatttg tgattagaca tgttataaac
      781 cactgcttta gtgactattt aaaacaatat attacatgtc gtaatcatgc aacctaacta
      841 tgttttcatt aatcaaatac aaagaataaa gagaaaagtg cgtagattca attatttggc
      901 atagactcaa aagagtgtat atatatctga cttttattaa attattaaac acaaatacat
      961 attttcataa gcaaaactat aaaagcccta aacatataac gattacctca aaggaaaaag
     1021 tcgttttctc ctaattaaaa gataggttac ttcctaatta atatataatt tatgtgaact
     1081 tcacaatata cagttcaata aaatttggta atttgaccga tttaaggaga gtggaaatta
     1141 gggcttctgc aatctttttt cttcgccgca atctcatgtc caattatcca ccgacggtgg
     1201 cggcgcaacc cacaacgacg gcgaatccac tgctgcagcg acatcaatct gaacagcgac
     1261 gaagagaatt accgaagatt gtcgaaacag agtctacaag tatggacatt acgatcggtc
     1321 aatctaagca gcctcaattt ttgaaatcca tagacgaatt agctgcgttt tcagttgcag
     1381 tggaaacatt caaacgccaa ttcgatgatc ttcagaagca catcgagtca atcgaaaacg
     1441 caattgattc caaactcgag agtaacggcg ttgtcctcgc cgcgcggaac aataatttcc
     1501 atcagccgat gttatcgcct ccgcggaaca atgtatctgt agaaaccacc gtcactgtga
     1561 gccaaccgtc tcaggagatt gtaccggaga cgtcgaataa accggagggg gaacgtatat
     1621 gtgagttgat gtgtagcaaa ggtctgcgta aatacatata cgcgaatatc tctgatcaag
     1681 ctaagttaat ggaagagatt ccttcagctt tgaaattggc caaggagcca gcgaagtttg
     1741 tattggattg tattggcaag ttttacttac aagggcgtag agcatttact aaagagtcgc
     1801 ctatgagctc tgcgagacaa gtttcgcttc ttatactgga gtcttttctt ctaatgcctg
     1861 atcgtggtaa agggaaggtg aagattgaga gttggattaa agatgaggcg gagacggctg
     1921 ctgttgcttg gaggaaaagg ttgatgactg aaggaggatt agctgcggct gagaaaatgg
     1981 atgcaagggg tttgctttta ctagttgctt gttttggtgt tccttcaaac tttaggagta
     2041 cagatttgct ggatttgata aggatgagtg gttcgaatga gattgccggt gctttgaagc
     2101 ggtcacagtt tcaggtacca tattctgttc tcactcggtg aatttcattg caaaggtggt
     2161 tccttttgtt gacatcatcg accaacatca agttccatct ttgtttttcg ataagcttga
     2221 tggtataaac taggagagca catcaaatat ttagagtgca atgactgatt gagccaaatc
     2281 ctagctagaa attaatctgg aaagaacttg gaactctcaa ccataggttt tggtacgaaa
     2341 ttgttgcttg tcagaaccaa atgataggct attgccttga aatagtgttt cttgtggttt
     2401 ccaatattgg aagttaaaat catatgactt agctgttgga tactaattaa gcttaagcaa
     2461 tgccaactct aagaagtggt acttacacaa tattctattg gtcataggta gctctagata
     2521 tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     2581 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
     2641 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     2701 tcactacttt ctcttatggt gttcaatgct tttcaagata cccagatcat atgaagcggc
     2761 acgacttctt caagagcgcc atgcctgagg gatacgtgca ggagaggacc atcttcttca
     2821 aggacgacgg gaactacaag acacgtgctg aagtcaagtt tgagggagac accctcgtca
     2881 acaggatcga gcttaaggga atcgatttca aggaggacgg aaacatcctc ggccacaagt
     2941 tggaatacaa ctacaactcc cacaacgtat acatcatggc cgacaagcaa aagaacggca
     3001 tcaaagccaa cttcaagacc cgccacaaca tcgaagacgg cggcgtgcaa ctcgctgatc
     3061 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     3121 tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
     3181 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaatagttga
     3241 atcaagtatc aagcgtggaa tgcatattga agctcttgaa atggtttata cctttggcat
     3301 ggaggataag ttttcagctg ctctagttct aacttcattc ttaaagatga gcaaggagtc
     3361 atttgagagg gcaaaacgga aagcccagtc accgctggca tttgtatgaa cccttccctt
     3421 gcacattatg tacctttatg aactctttat catcatctga gtctgaccat tgatatattt
     3481 atttctcaac agaaagaagc ggctacaaag cagctagctg tgttatcatc agttatgcag
     3541 tgtatggaga ctcacaagtt agatcctgcg aaagaactac caggatggca gatcaaagag
     3601 caaattgtta gcttggagaa agacactctt cagctcgaca aagagatgga agagaaagca
     3661 agatctctca gtttaatgga ggaagccgca cttgccaaga gaatgtataa ccaacagata
     3721 aaacgtccaa ggttatcacc catggaaatg ccaccagtaa cttcttcatc gtattctcct
     3781 atctaccgtg atagaagctt tcctagtcaa agagacgatg accaagatga aatatcagct
     3841 cttgtgagta gttacctcgg cccgtcaaca tcttttcctc atcgctcaag aagatccccg
     3901 gaatatatgg ttccacttcc acatggtggg ttaggaagaa gtgtatatgc atatgaacat
     3961 ctggccccaa attcatattc tccaggtcac ggacatagac ttcatcgaca gtactctccg
     4021 tctttggttc acggacagag acatccacta cagtactctc ctccaattca tggacaacaa
     4081 cagttaccat atggtataca aagggtttac agacattcac catctgaaga aagatatttg
     4141 ggtttatcca atcaaaggtc tcctcgcagt aactcatcat tagaccccaa ataggaggaa
     4201 tgtaaatttg taacaaagct ttttgttttt gcttaagtta gtcatttatt taactcccaa
     4261 cagtctcaaa atttaattta atgtttgggg cttaagaatg caaatttttt tgctcctgta
     4321 attgacattt aagatgctaa tgttattgct tcagaggttt tagtcaacct cagatacatc
     4381 gatatcacta tctaaataga cctctggctc ttggtcatct ggattctctt catcttctgt
     4441 ctctgttcct tcttgttctc gttgcactgc tcgagcaatt gcggattcca accttgtgct
     4501 tacagtttcc catgacacaa gcttttccat gaatgtattt atgtccgcct tcttatcttt
     4561 cttgaggaag atgaattcac cgaagatcca acttgagctt gacaatcaat caaatccgaa
     4621 acagaaacag agctttttga catctttgat ttagcagtct ttgatcttga ggaatatcaa
     4681 tgaacactag atacactcac acttgcaggc tttaaactgg attttaaaca tgaatagaag
     4741 cattgattcc atggaatgtg gtaagtgaca tagctggact tcttaaacaa atgtatgaac
     4801 gggtagggtt cattacaatg tagttataca gcactgagat ttatggaaga aaaaaaggac
     4861 acagctttaa ctatctacag agagacaaga acactaaaga caagagaatc ataagttcag
     4921 gagttcgtta aaatggctct attcaaatca cacattggca caagaccact aataagatac
     4981 caagtgggac aatcgaaaga taataagaga tagcatatca gagagagaga gagatttttt
     5041 gaggagggag aagttcgccg gaggcttctg tttctagtcg gcgttgtgat tggaattgtt
     5101 tgagcggtct ttccagtccc ggaattggaa acaagagggc taattttggt ccttttggaa
     5161 agatttgaaa cgacatcgtt attaagccct aattctacta taacgatgtc gtttacgaga
     5221 aacatacata tacatacttg agaataaagc ccattaccaa gagaccaatt ttggtctttt
     5281 tcgtatagta ttaactctac ggcgattgat ctaagcccaa taattgtgga acttgctcga
     5341 cttaggtctc aatctctatt ttcttgcagc ccgggggatc cactagttct agagcggccg
     5401 ccaccgcggt ggagctcacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     5461 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     5521 tccagctgca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     5581 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca gggggtgtta
     5641 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     5701 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     5761 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     5821 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     5881 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     5941 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     6001 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     6061 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     6121 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     6181 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     6241 accttatttt tgacgagggg aaattaggcc gggaagccga tctcggcttg aacgaattgt
     6301 taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt
     6361 gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca
     6421 ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc
     6481 cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac
     6541 ctgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc
     6601 gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg
     6661 gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat
     6721 ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac
     6781 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct
     6841 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     6901 catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
     6961 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
     7021 cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg
     7081 tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt cctggctggc
     7141 gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct tgctgttctt
     7201 ctacggcaag gtgctgtgca cggatctgcc ctggcttcag gagatcggaa gacctcggcc
     7261 gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc tcggttttct
     7321 ggaaggcgag catcgtttgt tcgcccagga ctctagctat agttctagtg gttggctact
     7381 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     7441 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     7501 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     7561 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     7621 cggcgcaagc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7681 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7741 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7801 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     7861 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7921 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7981 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8041 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8101 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8161 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8221 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8281 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8341 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8401 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     8461 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     8521 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     8581 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     8641 tggcagttcc ctactctcgc
//