LOCUS       sequence-466378-        7129 bp DNA     circular SYN 01-APR-2025
DEFINITION  AC5 promoter driven LIDAR reporter RNA for drosophila cell 
            expression (mCherry as marker, EGFP as output).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7129)
  AUTHORS   Zhang X, Mille-Fragoso LS, Kaseniit KE, Lee AP, Zhang M, Call CC, Hu
            Y, Xie Y, Gao XJ
  TITLE     Post-transcriptional modular synthetic receptors.
  JOURNAL   Nat Chem Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w.
  PUBMED    40155716
REFERENCE   2  (bases 1 to 7129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 1, 2025 from SnapGene Server 7.0.3
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w."
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      909..1497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1398..1417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2098..2232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2135..2154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2189..2208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2265..2282
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2305..2322)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2332..2373)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2412..3125)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(3132..3197)
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             complement(3366..3431)
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             complement(3477..4184)
                     /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(3752..3771)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4021..4039
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      4181..4190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4243..6752)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     primer_bind     complement(6816..6834)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6902..7006
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7007..7129,1..738)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
       61 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      121 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      421 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1021 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttgaag ctgtccctga
     1561 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     1621 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     1681 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     1741 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     1801 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     1861 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     1921 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     1981 atcggtcgac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2041 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2101 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2161 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2221 catgtctgga tcctcgatcg aggctgatca gcgagctcag gccttagaag gcacagtcga
     2281 ggctgatcag cgggtttaaa ctcaatggtg atggtgatga tgaccggtac gcgtagaatc
     2341 gagaccgagg agagggttag ggataggctt accttcgaac cgcgggccct ctagactcga
     2401 gcggccgcct acttatacag ttcgtccatc ccgagtgtta tcccagccgc agtgacgaac
     2461 tcaagaagga ccatgtgatc ccgtttttcg tttgggtcct tgctaagagc tgactgagta
     2521 gacaaatagt gattatcagg gagaagtact ggaccatccc ctatcggcgt gttctgttgg
     2581 taatggtccg caagctggac agagccgtcc tcaatattgt gtcggatttt aaagtttacc
     2641 ttgatgccgt ttttctgttt gtccgccatg atatagacat tatgagaatt atagttgtat
     2701 tcgagcttgt ggcccagtat attgccatcc tccttaaaat ctatgccctt cagttcgatc
     2761 cggttcacga gagtgtcacc ctcgaatttt acctcggctc tagttttgta gtttccgtcg
     2821 tctttaaaaa atatcgttcg ctcctgtacg taaccttcag gcatagcgga tttaaaaaaa
     2881 tcgtgctgtt tcatatggtc tggataccgg ctaaaacact gaacaccata tgtcagggtg
     2941 gtgacgagag taggccacgg cactggcaat tttccggtgg tacatatgaa cttcagcgta
     3001 agtttcccat atgtcgcgtc cccctcaccc tcaccactaa ctgagaattt atgcccatta
     3061 acgtccccgt ccaattcgac gaggatcgga acaacgcccg taaaaagttc ctcgcccttt
     3121 gagacactcg agggtcctgg attagattcc acatcacctg cgagtttgag tagatcgaag
     3181 ttgagtgttt gctttactgg ggctacgatc ttttgcttat gccgtgcctc actagtaccg
     3241 gtcgtctggg taggtgggat actatggtac gctgatgctg taccagacag accctgatgg
     3301 tgtctgagac gtaccctgat ggtgtacgtt actattccac ccacctaaac ggaggatccg
     3361 gtaccgggac cggggttgga ctcgacgtct cccgccagct tcagaaggtc aaaattcaaa
     3421 gtctgtttca ccggagccac aattttctgt ttgtgtctgg cttccaacgc gtcgctcttg
     3481 tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg
     3541 atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg
     3601 ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc gtagtggccg
     3661 ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac
     3721 atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag
     3781 gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag
     3841 tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag
     3901 ccctcgggga aggacagctt caagtagtcg ggaatgtcgg cggggtgctt cacgtaggcc
     3961 ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc
     4021 ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc
     4081 tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac
     4141 tccttgatga tggccatgtt atcctcctcg cccttgctca ccatggtggc gatcccattt
     4201 cggaattcca ccacactgga ctagtaggta ccccgatccg gggtctctgg attagacgac
     4261 tgctggctga tggagcggct ttgtgtcggg aggagtatcc acacagcaca agaactcaaa
     4321 cggtagtgat atgaaaactg gtcccgcagc ccgttttata aagcccgctt agcgctaaaa
     4381 ccttatcagc cgcaatgctc tccgctctca agtcgcgttc aaaactttta ccatatatgg
     4441 gtatgaattg ttcgccgatt gggttttcat ttacggagaa tttcctccgc aactgggtgt
     4501 tcgtgcttgt gtgtgtcttg tatatcgcca tgaagagtgt acttccatgg gtcctatcag
     4561 ttgcggcatg tgattcaatt tgggggttac tcatattcga tacttcagct ggttatcttt
     4621 atgctgtgat ggtcacggtg cttttacaca ttttttcccc aactactcat tgtatgccaa
     4681 aacatacata tgtacatgtg tttatagtgg gtttattagt aatgtccagc gatattaaaa
     4741 cattacagtt aaataatatt catattagct tacgtttact gctatctgca cttcaaattg
     4801 gttattccag ggacattttg tgcaacacac atcttctagt agggcattca ctagtcgtag
     4861 aaaacacacc taacttaagc cggttttaac aataacggcg aaacagctgc cagacgagag
     4921 aaaagtacac agcaagaagc atttctataa tgatgccgtc ctaaatatgc agcatttttg
     4981 ctcaaattga tcatttcgtt gcatcgctca ggtggtatag tatttggcta ttgaatgtga
     5041 ttgctatctt aatagttttg aatagtaata gtgcaaatat tcattcataa aaaataggta
     5101 atcgattttt tcagtgcagt ctatgcgttt gcgcaacaga gacgaagaga gagacgcctc
     5161 acacacaagc acatgcggtc taagctgctc tccctctttt tctctctctc tctggcttgc
     5221 ggcagttttc cagaggctgt gactcagtaa gcatttcttg ttgttaccct aacgataagc
     5281 gtccgattcc taattctctt tcaaataatt tgttttattt ttttttttgt gtgttttgaa
     5341 gtcaaacagc ttcataaaca atattaaata tcgcaattgt tattgcaaat gcattcgtat
     5401 caggcgcctt gatgtttttc gccaaaactt cgtctgctct ttgtcagcgt aattgcgttt
     5461 gccggtcgtg ccgtcttgct cgcacgcggc ttgcaaacaa aaagtgaaag agggaaagcc
     5521 acctgtgttg ttgcttttgt tgctatcgtt ggctgcacac acatggcaat aagtaaccat
     5581 attgtagact tatttatttt agcgttttcg ttgactaatc tttgtttaaa taattgccta
     5641 tttcgagcca aaatgacgca taaaaggaaa cggcaacatg cggtgcgttg agcatgcgta
     5701 agcgagatgg cgctagagag agcgcagcat aaaaagaaaa aaacaaccgg cgagtggcga
     5761 gacacgaata atacacacac gcacacatgg tcaaaggagt ggctccaacg tcggtgacgt
     5821 cgcgcccagc cacagacccc gccacgcccc gccccaccct tccttccgct gccccgcacc
     5881 cctttccttc cactaccgtt ttgccgcatt tggtattgcc atctcgcttt tttggcttta
     5941 ctttgctttt tgaataagcc aagaaactga aatcaaagcg gacagtccga aaattgaata
     6001 ttccaaaaaa caacaattgc acatgcactg cgttcggcaa aagacgcaga aattcaagcc
     6061 ttttggtctg gcatcatctt aaaatatcat cagtatcatc attgccatta taaatacata
     6121 agaaaatgag aatacattaa aggtatatac atacaatctt tgtcatagtc aacatatttt
     6181 gcaacaaaat tatctataga taacatgttt ttgtagaaaa ttccactaac tgattttcgt
     6241 ataaatacaa attattctca ttcttcaaaa tactcgccga cgtcatcgcg aaattcctca
     6301 gcattcttcg acaaatgaag agagacagcg agtgacacaa tcttatcttg ccgacgagtc
     6361 tttgaaaatg gccccgggta acggtgaaaa acatatagcc aagaatggag cgatcgccag
     6421 gcgatggaaa ttggcttcaa attgctccga atatggccag ttggtcagtt ggcattggat
     6481 tgtggagaga tactagatca gctcgttctc cttctcctcc gcattctcct actccgcccg
     6541 aagacgcttc tttgatcgct tctttctcga atgtgtacat attcattatg tatttggtgt
     6601 ttggcgacga tcagccagct ttaatggcca taaaacgtgt cttggtttgt gggccacata
     6661 acaggcagcg agatctggaa aaatgatgtg acagtggaaa tgagaagtag gtgcatctgc
     6721 aaagatttga ttcagagttg atgccattca tgattttttt ttagaagtat catttgtgtt
     6781 tttagaatat agaattaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     6841 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6901 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6961 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7021 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7081 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacga
//
