LOCUS       sequence-479168-        5735 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  mScarlet-I3 expressed under a glial specific promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5735)
  TITLE     Fyhn unpublished plasmids
REFERENCE   2  (bases 1 to 5735)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV gfaABC1D:mScarlet-I3
FEATURES             Location/Qualifiers
     source          1..5735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     447..466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    834..855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(870..875,876..893,894..900)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    908..924
                     /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     913..935
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     932..948
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     932..948
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   986..1115
                     /label=AAV2 ITR
     intron          1966..2098
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2081..2100
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     2143..2162
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2143..2161
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2168..2177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2174..2860
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGGT
                     VSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     complement(2304..2322)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    2881..3469
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2934..2954)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3352..3363)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3487..3608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3498..3517)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3552..3571
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3770..3899
                     /label=AAV2 ITR
     primer_bind     complement(3918..3935)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3918..3934)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3927..3949)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4076..4531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4163..4182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4373..4394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4557..4661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4662..4730,4731..5522)
                     /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"
     rep_origin      join(5693..5735,1..546)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
       61 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      121 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      181 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      241 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      301 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      361 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      421 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      481 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      541 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      601 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      661 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      721 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      781 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      841 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      901 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
      961 agatttaatt aaggccttaa ttaggctgcg cgctcgctcg ctcactgagg ccgcccgggc
     1021 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     1081 agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg
     1141 ctacttatct acgtagccat gctctaggaa gatctaacat atcctggtgt ggagtagggg
     1201 acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg gaggaggcag
     1261 acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc ccccagggcc
     1321 tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg gggtgggcac
     1381 agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa gcccattgag
     1441 cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa aagcagcaca
     1501 gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca aggctctatt
     1561 cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg ggtggcaggg
     1621 ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt gaggggagag
     1681 ctctccccat agctgggctg cggcccaacc ccaccccctc aggctatgcc agggggtgtt
     1741 gccaggggca cccgggcatc gccagtctag cccactcctt cataaagccc tcgcatccca
     1801 ggagcgagca gagccagagc aggttggaga ggagacgcat cacctccgct gctcgcaagc
     1861 tttattgcgg tagtttatca cagttaaatt gctaacgcag tcagtgcttc tgacacaaca
     1921 gtctcgaact taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
     1981 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     2041 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     2101 gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag
     2161 ggctagcgcc accatggata gcaccgaggc agtgatcaag gagttcatgc ggttcaaggt
     2221 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggacg
     2281 accctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     2341 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc
     2401 cgccgacatc cccgactact ggaagcagtc cttccccgag ggcttcaagt gggagcgcgt
     2461 gatgatcttc gaggacggcg gcaccgtgtc cgtgacccag gacacctccc tggaggacgg
     2521 caccctgatc tacaaggtga agctccgcgg cggcaacttc cctcctgacg gccccgtaat
     2581 gcagaagagg acaatgggct gggaagcatc caccgagcgg ttgtaccccg aggacgtcgt
     2641 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga
     2701 cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct tcaacatcga
     2761 ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg
     2821 ctccgtggcc cgccactcca ccggaggctc aggtggctcc taaggcgcgc ccctgaattc
     2881 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2941 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3001 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3061 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3121 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3181 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3241 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3301 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3361 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3421 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg cttcgagcag
     3481 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3541 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3601 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     3661 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     3721 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3781 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3841 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3901 cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3961 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4021 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     4081 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4141 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4201 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4261 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4321 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4381 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4441 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4501 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5701 ctttttttct gcgcgtaatc tgctgcttgc aaaca
//
