LOCUS       sequence-81956-e        6904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of TagRFP-tagged PSD-95_Y12E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6904)
  TITLE     Hell lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6904)
  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            "Unpublished"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: PSD-95_Y12E-pTagRFP
FEATURES             Location/Qualifiers
     source          1..6904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      621..630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2844..2853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2850..3563
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3722..3741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3776..3795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3813..4268)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3950..3971)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4162..4181
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4295..4399
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4401..4758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4401..4592
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4424..4444)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4609..4744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4793..5587
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4793..5587
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4847..4866)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5457..5476
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5775..5794)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5819..5866
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6195..6783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6684..6703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatctcga gctcaagctt gccaccatgg actgtctctg tatagtgaca accaagaaag
      661 agcgctacca agatgaagac acgccccctc tggaacacag cccggcccac ctccccaacc
      721 aggccaattc tccccctgtg attgtcaaca cggacaccct agaagcccca ggatatgagt
      781 tgcaggtgaa tggaacagag ggggagatgg agtatgagga gatcacattg gaaaggggta
      841 actcaggtct gggcttcagc atcgcaggtg gcactgacaa cccgcacatc ggtgacgacc
      901 cgtccatttt tatcaccaag atcattcctg gtggggctgc agcccaggat ggccgcctca
      961 gggtcaatga cagcatcctg tttgtaaatg aagtggatgt tcgggaggtg acccattcag
     1021 ctgcggtgga ggccctcaaa gaggcaggtt ccatcgttcg cctctatgtc atgcgccgga
     1081 aacccccagc cgaaaaggtc atggagatca aactcatcaa agggcctaaa ggacttggct
     1141 tcagcattgc ggggggcgtt gggaaccagc acatccctgg agataacagc atctatgtaa
     1201 cgaagatcat cgaaggaggt gctgcccaca aggatggcag gttgcagatt ggagacaaga
     1261 tcctggcggt caacagtgtg gggctggagg acgtcatgca cgaggatgcc gtggcagccc
     1321 tgaagaacac atatgacgtt gtgtacctaa aggtggccaa gcccagcaat gcctacctga
     1381 gtgacagcta tgctccccca gacatcacaa cctcgtattc tcagcacctg gacaatgaga
     1441 tcagtcatag cagctacttg ggcactgact accccacagc catgaccccc acttcccctc
     1501 ggcgctactc ccctgtggcc aaggacctgc tgggggagga agacattccc cgggaaccaa
     1561 ggcggatcgt gatccatcgg ggctccaccg gcctgggctt caacatcgtg ggcggcgagg
     1621 atggtgaagg catcttcatc tccttcatcc ttgctggggg tccagccgac ctcagtgggg
     1681 agctacggaa gggggaccag atcctgtcgg tcaatggtgt tgacctccgc aatgccagtc
     1741 acgaacaggc tgccattgcc ctgaagaatg cgggtcagac ggtcacgatc atcgctcagt
     1801 ataaaccaga agagtatagt cgattcgagg ccaagatcca tgatcttcgg gaacagctca
     1861 tgaatagtag cctaggctca gggactgcat ccttgcgaag caaccccaag aggggcttct
     1921 acattagggc cctgtttgat tacgacaaga ccaaggactg cggtttcttg agccaggccc
     1981 tgagcttccg cttcggggat gtgcttcatg tcattgacgc tggtgacgaa gagtggtggc
     2041 aagcacggcg ggtccactcc gacagtgaga ccgacgacat tggcttcatt cccagcaaac
     2101 ggcgggtcga gcgacgagag tggtcaaggt taaaggccaa ggactggggc tccagctctg
     2161 gatcacaggg tcgagaagac tcggttctga gctatgagac ggtgacccag atggaagtgc
     2221 actatgctcg tcccatcatc atccttggac ccaccaaaga ccgtgccaac gatgatcttc
     2281 tctccgagtt ccccgacaag tttggatcct gtgtccctca tacgacacgt cctaagcggg
     2341 aatatgagat agacggccgg gattaccact ttgtctcctc ccgggagaaa atggagaagg
     2401 acatccaggc acacaagttc attgaggctg gccagtacaa cagccacctc tatgggacca
     2461 gcgtccagtc tgtgcgagag gtagcagagc aggggaagca ctgcatcctc gatgtctcgg
     2521 ccaatgccgt gcggcggctg caggcggccc acctgcaccc catcgccatc ttcatccgtc
     2581 cccgctccct ggagaatgtg ctagagatca ataagcggat cacagaggag caagcccgga
     2641 aagccttcga cagagccacg aagctggagc aggagttcac agagtgcttc tcagccatcg
     2701 tagagggcga cagctttgaa gagatctatc acaaagtgaa acgtgtcatt gaagacctct
     2761 caggccccta catctgggtc ccagcccgag agagactcgg aattctgcag tcgacggtac
     2821 cgcgggcccg ggatccaccg gtcgccacca tggtgtctaa gggcgaagag ctgattaagg
     2881 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
     2941 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
     3001 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
     3061 gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg
     3121 gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg
     3181 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
     3241 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
     3301 tgtaccccgc tgacggcggc ctggaaggca gaagcgacat ggccctgaag ctcgtgggcg
     3361 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
     3421 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
     3481 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     3541 aactggggca caaacttaat tgagcggccg cgactctaga tcataatcag ccataccaca
     3601 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3661 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3781 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3841 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3901 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3961 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4021 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4081 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4141 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4201 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4261 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4321 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4381 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4441 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4501 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4561 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4621 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4681 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4741 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4801 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4861 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4921 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4981 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5041 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5101 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5161 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5221 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5281 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5341 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5401 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5461 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5521 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5581 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5641 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5701 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5761 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5821 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5881 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5941 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6001 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     6901 ttat
//
