LOCUS       sequence-371426-        6891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red Fluorescent Protein for Long-Term Labeling of Fine Structures in
            Neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6891)
  AUTHORS   Ning L, Geng Y, Lovett-Barron M, Niu X, Deng M, Wang L, Ataie N, 
            Sens A, Ng HL, Chen S, Deisseroth K, Lin MZ, Chu J
  TITLE     A Bright, Nontoxic, and Non-aggregating red Fluorescent Protein for 
            Long-Term Labeling of Fine Structures in Neurons.
  JOURNAL   Front Cell Dev Biol. 2022 Jun 29;10:893468. doi: 
            10.3389/fcell.2022.893468. eCollection 2022.
  PUBMED    35846353
REFERENCE   2  (bases 1 to 6891)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Cell 
            Dev Biol."; date: "2022-06-29"; volume: "10:893468. doi"; pages: " 
            10.3389/fcell.2022.893468. eCollection 2022"
COMMENT     Sequence Label: pC3.1-caggs-Crimson-CAAX
FEATURES             Location/Qualifiers
     source          1..6891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1321..1920)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1404..1424)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1901..1920
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1931..2260)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1945..2080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1999..2018)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2245..2265
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2274..2702)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2384..2405)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2596..2615
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2748..2972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2961..2978
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     regulatory      3880..3889
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4022..4041)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          4049..5065
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4103..4125)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(5066..5343)
                     /label=chicken beta-actin promoter
     enhancer        5345..5724
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5756..5860
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5861..5929,5930..6721)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6079..6098)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa
     1321 tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc
     1381 ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc
     1441 caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct
     1501 gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg
     1561 ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga
     1621 accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct
     1681 ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat
     1741 gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg
     1801 gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
     1861 tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
     1921 gcgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     1981 gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     2041 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     2101 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     2161 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     2221 ctggggagcc tggggacttt ccacacccta actgacacac attccacaga attaattcgc
     2281 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2341 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2401 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2461 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2521 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2581 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2641 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2701 gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac
     2761 cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
     2821 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
     2881 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
     2941 aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg tttaaacggg
     3001 ccctctagac tcgagcggcc gccactgtgc tggatatctg cagaattcca ccacactgga
     3061 ctagtggatc cgagctcggt accaagcttc attacatgat cacgcactta gtcttggact
     3121 tcttcttttt cttcttgccg tccttgctca tcttcttaag cttgtacagc tcgtccatgc
     3181 cattaagttt gtgccccagt ttgctaggga ggtcgcagta tctggccaca gccacctcgt
     3241 gctgctcgac gtaggtctcg ttgtcggcct ccttgattct ttccagtctg cggtccacgt
     3301 agtggacgcc gggcatcttg acgggtttct tggatttgta tgtggtcttg aagttgcagt
     3361 gcaggtggcc cccgcccacg agcttcaggg ccatgtcgca tctgccttcc agggcgccgt
     3421 cagcggggta cagggtctcg gtgctgggct cccagcccag tgttgtctgc ttcatcacgg
     3481 ggccgttggc tgggaagttc acccctctca gcttcacgtt gtagatgagc tcgccgtcct
     3541 ggaggctggt gtcctgggtg gcggtcagca cgcccccgtc ttcgaacacc atggttctct
     3601 cccatgtgaa gccctcaggg aaggactgct taaaataatc ggggaggccc ttggggtact
     3661 tgatgaaggc cttgctcccg tacataaaca tggtggccag gatgtcgaat gcgaacggca
     3721 gggggcctcc ctccaccacc ttgatcctgt tggtctgggt gccctcgtag ggcttgccct
     3781 ccccttcgtg ggtgcacttg aactggtggc cgttcacgct gccttccagg tacagcttgc
     3841 ttctcatgtt ctccttgatc agctcctcgc ccttgctcac catggtggcg gctagctcat
     3901 gcagatctca gtggtatttg tgagccaggg cattggccac accagccacc accttctgat
     3961 aggcagcctg cacctgagga gtgaattcct cgaggaattc tttgccaaaa tgatgagaca
     4021 gcacaataac cagcacgttg cccaggagct gtaggaaaaa gaagaaggca tgaacatggt
     4081 tagcagaggc tctagagccg ccggtcacac gccagaagcc gaaccccgcc ctgccccgtc
     4141 ccccccgaag gcagccgtcc ccccgcggac agccccgagg ctggagaggg agaaggggac
     4201 ggcggcgcgg cgacgcacga aggccctccc cgcccatttc cttcctgccg gcgccgcacc
     4261 gcttcgcccc gcgcccgcta gagggggtgc ggcggcgcct cccagatttc ggctccgcac
     4321 agatttggga caaaggaagt ccctgcgccc tctcgcacga ttaccataaa aggcaatggc
     4381 tgcggctcgc cgcgcctcga cagccgccgg cgctccgggg gccgccgcgc cccycccccg
     4441 agcccycccc ggcccgaggc ggccccgccc cgcccggcac ccccacctgc cgccaccccc
     4501 cgcccggcac ggcgagcccc gcgccacgcc ccgtacggag ccccgcaccc gaagccgggc
     4561 cgtgctcagc aactcgggga ggggggtgca gggggggttg cagcccgacc gacgcgccca
     4621 caccccctgc tcaccccccc acgcacacac cccgcacgca gcctttgttc ccctcgcagc
     4681 cccccccgca ccgcggggca ccgcccccgg ccgcgctccc ctcgcgcaca ctgcggagcg
     4741 cacaaagccc cgcgccgcgc ccgcagcgct cacagccgcc gggcagcgcg gagccgcacg
     4801 cggcgctccc cacgcacaca cacacgcacg caccccccga gccgctcccc ccgcacaaag
     4861 ggccctcccg gagcccctca aggctttcac gcagccacag aaaagaaaca agccgtcatt
     4921 aaaccaagcg ctaattacag cccggaggag aagggccgtc ccgcccgctc acctgtggga
     4981 gtaacgcggt cagtcagagc cggggcgggc ggcgcgaggc ggcggcggag cggggcacgg
     5041 ggcgaaggca gcgcgcagcg actcccgccc gccgcgcgct tcgcttttta tagggccgcc
     5101 gccgccgccg cctcgccata aaaggaaact ttcggagcgc gccgctctga ttggctgccg
     5161 ccgcacctct ccgcctcgcc ccgccccgcc ccycsccccs ccccsccccs ccyggcgcgc
     5221 gccccccccc cccccccgcc cccatcgctg cacaaaataa ttaaaaaata aataaataca
     5281 aaattggggg tggggagggg ggggagatgg ggagagtgaa gcagaacgtg gggctcacct
     5341 cgaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     5401 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     5461 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     5521 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     5581 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     5641 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     5701 aataactagt caataatcaa tgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg
     5761 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5821 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5881 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5941 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6001 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6061 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6121 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6181 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6241 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6301 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6361 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6421 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6481 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6541 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6601 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6661 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6721 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6781 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6841 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
