AttB-miRFP680-P2A-PAmCherry-NLS-EGFP-3xNLS

LOCUS       AttB-miRFP680-P2A-PAmCherry-NLS-EGFP-3xNLS             5773 bp DNA     circular SYN 03-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5773)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 1, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..64)
                     /label=M13-rev
     misc_binding    complement(70..92)
                     /label=LacO
     rep_origin      complement(432..1114)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(1212..1871)
                     /label=AmpR
     rep_origin      complement(2332..2638)
                     /direction=LEFT
                     /label=F1 ori
     CDS             complement(2657..2725)
                     /label=LacZ alpha
     primer_bind     2796..2813
                     /label=M13-fwd
     primer_bind     2812..2839
                     /label=T7
     misc_feature    2910..2930
                     /label=attB-Left
     misc_feature    2931..2951
                     /label=attB-Right
     misc_feature    2971..2979
                     /label=Feature 13
     misc_feature    2974..2979
                     /label=Kozak sequence
     CDS             2980..3927
                     /label=miRFP680
     primer_bind     2980..2998
     misc_feature    complement(3909..3927)
                     /label=Geneious type: primer_bind_reverse
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    3928..3993
                     /label=P2A
                     /note="Geneious type: motif"
     primer_bind     3928..3946
     misc_feature    3980..3994
                     /label=Gibson
     primer_bind     3984..3999
     misc_structure  4000..4707
                     /label=PAmCherry1
     misc_structure  4714..4734
                     /label=SV40-NLS
     misc_feature    join(4744..4938,4948..5460)
                     /label=eGFP
                     /label=eGFP(1)
     misc_feature    4939..4947
                     /label=eGFP Chromophore
     misc_feature    4939..4947
                     /label=eGFP
     CDS             5467..5487
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5491..5511
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5515..5535
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5545..5773
                     /label=SV40_term
ORIGIN
        1 ggagatcccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt
       61 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      121 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      181 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      241 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      301 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      361 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
      901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
      961 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1021 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1081 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1141 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1201 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1261 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1321 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1381 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1441 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1501 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1561 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1621 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1681 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     1741 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     1801 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     1861 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     1921 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     1981 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2041 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2101 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2161 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     2221 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2281 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2341 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2401 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     2461 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     2521 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     2581 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     2641 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2701 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2761 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     2821 aatacgactc actatagggc gaattgggta ccgggccccc cctcgaggtc gacgatgtag
     2881 gtcacggcat tccggactca gcgctcgagc cggcttgtcg acgacggcgg tctccgtcgt
     2941 caggatcatc cgcagattat accgcaacta cacgccacca tggcggaggg cagcgtggcg
     3001 cgtcagccgg atctgctgac ctgtgatgat gaaccgatcc acattccggg cgcgatccag
     3061 ccgcacggtc tgctgctggc gctggcagcg gatatgacca tcgtggcggg ctctgataac
     3121 ctgccggaac tgaccggcct ggcaatcggt gcgctgatcg gtcgttccgc ggcggatgtt
     3181 ttcgattctg aaacccataa ccgtctgacc atcgcgctgg cggaaccggg cgcggcggtt
     3241 ggtgcgccga tcaccgttgg cttcaccatg cgtaaagacg cgggctttat cggtagctgg
     3301 caccgtcacg atcagctgat ctttctggaa ctggaaccgc cgcagcgcga tgtggcggaa
     3361 ccgcaagcgt tcttccgtcg taccaactct gcgattcgtc gtctgcaagc ggcggaaacc
     3421 ctggaaagcg cgtgcgcggc tgcggcgcag gaagttcgta aaatcaccgg cttcgatcgt
     3481 gttatgatct atcgtttcgc gtctgacttc tccggcgaag tgatcgcgga agatcgttgc
     3541 gcagaagttg aatctaaact gggtctgcac tatccggcgt ccaccgtccc ggcccaggcg
     3601 cgccgtctgt ataccatcaa cccggttcgt attatcccgg atattaacta tcgtccggtt
     3661 ccagtaaccc cggatttaaa cccggtgacc ggtcgtccga ttgatctgag ctttgctatc
     3721 ctgcgttctg ttagcccgtg ccacctggaa tttatgcgta acatcggtat gcatggtacc
     3781 atgagcatct ctatcctgcg tggtgaacgt ctgtggggtc tgatcgtttg ccaccaccgt
     3841 actccgtact atgttgacct ggacgggcgc caggcatgca aacgtgtagc tgagcgtctg
     3901 gccacccaaa tcggtgttat ggaggaaggc tctggagcca ccaacttcag cctgctgaag
     3961 caggcaggcg acgtggaaga gaaccctggc cctggatcca tggtgagcaa gggcgaggag
     4021 gataacatgg ccatcattaa ggagttcatg cgcttcaagg tgcacatgga ggggtccgtg
     4081 aacggccacg tgttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     4141 accgccaagc tgaaggtgac caagggcggc cccctgccct tcacctggga catcctgagc
     4201 cctcagttca tgtacggctc caatgcctac gtgaagcacc ccgccgacat ccccgactac
     4261 tttaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaaatt cgaggacggc
     4321 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     4381 aagctgcgcg gcaccaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc
     4441 tgggaggccc tctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgaggtcaag
     4501 cccagagtga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     4561 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca accgcaagct ggacatcacc
     4621 agccacaacg aggactacac catcgtggag cagtacgaga gagccgaggg ccgccactcc
     4681 accggcggca tggacgagct gtacaagtct ggccctaaga aaaagcggaa agtgggaggc
     4741 agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4801 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4861 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4921 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4981 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     5041 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5101 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     5161 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     5221 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     5281 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     5341 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     5401 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5461 acgcgtccaa aaaagaagag aaaggtagac ccgaagaaaa agcgtaaggt cgatcccaaa
     5521 aagaaacgga aagtgtaagg atcctgatca taatcagcca taccacattt gtagaggttt
     5581 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5641 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5701 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5761 tcaatgtatc tta
//