LOCUS       sequence-250183-        6307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Labeling of cre-expressing neurons (red fluorescence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6307)
  AUTHORS   Bensussen S, Shankar S, Ching KH, Zemel D, Ta TL, Mount RA, Shroff 
            SN, Gritton HJ, Fabris P, Vanbenschoten H, Beck C, Man HY, Han X
  TITLE     A Viral Toolbox of Genetically Encoded Fluorescent Synaptic Tags.
  JOURNAL   iScience. 2020 Jun 30;23(7):101330. doi: 10.1016/j.isci.2020.101330.
  PUBMED    32674057
REFERENCE   2  (bases 1 to 6307)
  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: "iScience.";
            date: "2020-06-30"; volume: "23(7):101330. doi"; pages: " 
            10.1016/j.isci.2020.101330"
COMMENT     Sequence Label: AAV-EF1A-DIO-mRuby2
FEATURES             Location/Qualifiers
     source          1..6307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   294..423
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        524..1702
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          755..1693
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1650..1670
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1733..1766)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1817..1850)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1862..2572)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     regulatory      2569..2578
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2585..2618
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2669..2702
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2727..3315
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2780..2800)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3198..3209)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3347..3823
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3863..4003
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3863..3992
                     /label=AAV2 ITR
     rep_origin      4078..4533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4165..4184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4375..4396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4550..4569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4669..4691
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4729..4747)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4815..4919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4920..4988,4989..5780)
                     /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(5138..5157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5951..6307,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
      301 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
      361 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      421 cctgcggccg cacgcgtaag ctttgcaaag atggataaag ttttaaacag agaggaatct
      481 ttgcagctaa tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca
      541 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      601 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
      661 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
      721 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
      781 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
      841 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
      901 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
      961 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     1021 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     1081 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     1141 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     1201 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     1261 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     1321 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     1381 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     1441 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     1501 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     1561 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     1621 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     1681 tcttccattt caggtgtcgt gaggtaccgg atcctctaga gtcgactccg gaataacttc
     1741 gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc tgctattagc
     1801 aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat ggcgcgcctt
     1861 acttgtacag ctcgtccatc ccaccaccaa gcccggcgaa cttggcaact gcgtgttcgc
     1921 gttgtactac gaacatttca ttgtcacttt cctctaacct ttccaggcgg tgatcaacgg
     1981 catggatacc gggcatcttg atgttcccga cggtcttttt tgacctgtaa gttgttacga
     2041 aagagcaaga cagatggcca ccaccatcaa ctttcagtgc catatgagtg tatcccctca
     2101 gaccaccatc tgctggatac atcatctctg tattaggctc ccaacccttg gtcttcttct
     2161 gcatcacggg accattggag ggaaagttta cccctctgac ttggacgtgg taaacgagac
     2221 agccatcctc aaggctggtg tcctgcatga cggtgacgac tccaccatct tcgtatctcg
     2281 taactctttc ccaagtaaaa ccctcaggaa aggactgttt aaagaaatca ggaatgcctt
     2341 tcgggtactt gataaaagta cggctgccat acatgaacga cgtggcaaga atgtcaaagg
     2401 caaatggcag gggtcctccc tcgatgactt tgatcctcat ggtttgagtt cccatgtacg
     2461 gattgccttc tccttcacct gtgcatttga attggtggcc gttgaccgaa ccttccatga
     2521 ccaccttcat acgcatattt tccttgatca gctcttcgcc cttagacacc atggtggcgc
     2581 tagcataact tcgtataaag tatcctatac gaagttattt gccttaaccc agaaattatc
     2641 actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct atacgaagtt
     2701 atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2761 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2821 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2881 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2941 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3001 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3061 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3121 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3181 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3241 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3301 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3361 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3421 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3481 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3541 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3601 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3661 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3721 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3781 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     3841 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3901 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3961 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4021 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4081 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4141 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4201 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4261 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4321 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4381 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4441 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4501 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4561 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4621 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4681 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4741 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4801 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4861 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4921 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4981 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5041 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5101 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5161 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5221 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5281 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5341 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5401 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5461 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5521 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5581 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5641 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5701 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5761 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5821 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5881 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6061 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6301 agcgaac
//
