LOCUS       sequence-261221-        6042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to express mRuby3. Can also be used to create 
            adeno-associated virus for delivery of the mRuby3 sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6042)
  AUTHORS   Rylan Larsen
  TITLE     AAV red fluorescent protein plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6042)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AAV Ef1a-mRuby3-WPRE
FEATURES             Location/Qualifiers
     source          1..6042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1198..1908
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     misc_feature    1939..2527
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1992..2012)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2410..2421)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2559..3035
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2758..2777)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3075..3215
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3075..3204
                     /label=AAV2 ITR
     rep_origin      3290..3745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3377..3396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3587..3608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3762..3781)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3881..3903
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3941..3959)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4027..4131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4132..4200,4201..4992)
                     /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(4350..4369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5163..5751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5652..5671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5813..5942
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cggtggcatg
     1201 gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt catggaaggt
     1261 tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcagacc gtacgaggga
     1321 gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc ctttgacatt
     1381 cttgccacgt cgttcatgta tggcagccgt acctttatca agtacccggc cgacatccct
     1441 gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac gagatacgaa
     1501 gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga gctcgtctac
     1561 aacgtcaagg tcagaggggt aaactttccc tccaatggtc ccgtgatgca gaagaagacc
     1621 aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct gagaggatac
     1681 actgacatcg cactgaaagt tgatggtggt ggccatctgc actgcaactt cgtgacaact
     1741 tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc cgttgatcac
     1801 cgcctggaaa ggatcgagga gagtgacaat gaaacctacg tagtgcaaag agaagtggca
     1861 gttgccaaat acagcaacct tggtggtggc atggacgagc tgtacaagta ataggaattc
     1921 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1981 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2041 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2101 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2161 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2221 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2281 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2341 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     2401 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2461 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2521 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     2581 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     2641 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     2701 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     2761 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     2821 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2881 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2941 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     3001 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     3061 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3121 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     3181 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     3241 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     3301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3361 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3481 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     3541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3601 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     3661 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3721 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     3781 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3841 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3901 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3961 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4021 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4081 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4141 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4201 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4261 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4321 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4381 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4441 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4501 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4561 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4621 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4681 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4741 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4801 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4861 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4921 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4981 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5041 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5101 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5161 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5221 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5281 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5341 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5401 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5461 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5521 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5581 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5641 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5701 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5761 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
     5821 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
     5881 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     5941 ctgcggccgc acgcgtaagc tttgcaaaga tggataaagt tttaaacaga gaggaatctt
     6001 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa tt
//
