LOCUS       sequence-394752-        6816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For labeling pre-synaptic terminals.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  TITLE     Pre-synaptic marker
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6816)
  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            "Unpublished"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: CMV mRuby3-Synapsin1a
FEATURES             Location/Qualifiers
     source          1..6816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1323
                     /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"
     polyA_signal    3604..3725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3641..3660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3695..3714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3732..4187)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3869..3890)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4081..4100
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4214..4318
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4320..4677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4320..4511
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4343..4363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4528..4663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4712..5506
                     /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             4712..5506
                     /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(4766..4785)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5376..5395
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5694..5713)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5738..5785
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6114..6702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6603..6622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgtc taagggcgaa gagctgatca aggaaaatat gcgtatgaag
      661 gtggtcatgg aaggttcggt caacggccac caattcaaat gcacaggtga aggagaaggc
      721 agaccgtacg agggagtgca aaccatgagg atcaaagtca tcgagggagg acccctgcca
      781 tttgcctttg acattcttgc cacgtcgttc atgtatggca gccgtacctt tatcaagtac
      841 ccggccgaca tccctgattt ctttaaacag tcctttcctg agggttttac ttgggaaaga
      901 gttacgagat acgaagatgg tggagtcgtc accgtcacgc aggacaccag ccttgaggat
      961 ggcgagctcg tctacaacgt caaggtcaga ggggtaaact ttccctccaa tggtcccgtg
     1021 atgcagaaga agaccaaggg ttgggagcct aatacagaga tgatgtatcc agcagatggt
     1081 ggtctgagag gatacactga catcgcactg aaagttgatg gtggtggcca tctgcactgc
     1141 aacttcgtga caacttacag gtcaaaaaag accgtcggga acatcaagat gcccggtgtc
     1201 catgccgttg atcaccgcct ggaaaggatc gaggagagtg acaatgaaac ctacgtagtg
     1261 caaagagaag tggcagttgc caaatacagc aaccttggtg gtggcatgga cgagctgtac
     1321 aagtccggac tcagatctat gaactacctg cggcgccgcc tgtcggacag caacttcatg
     1381 gccaatctgc ctaatgggta tatgacagac ctgcagcgcc cgcaaccacc tccgccgccg
     1441 ccctcagccg ccagccctgg ggccactccc ggatccgctg ctgcctctgc cgagagggcc
     1501 tccacagctg ctccagtggc ctctccagca gcccctagtc ccgggtcctc ggggggcggt
     1561 ggcttcttct cctcgctgtc taacgcggtc aaacagacca cagcagctgc agccgccacc
     1621 ttcagtgagc aggtgggcgg tggctctggg ggcgcaggcc gcgggggcgc cgccgccagg
     1681 gtgctgctgg tcatcgacga gccgcacacc gactgggcaa aatacttcaa agggaagaag
     1741 atccatggag aaattgacat taaagtagag caagctgaat tctccgatct caatcttgtg
     1801 gctcatgcca atggtggatt ctccgtggac atggaagttc ttcggaatgg ggtcaaagtt
     1861 gtgaggtctc tgaagccaga ctttgtgctg atccgccagc atgccttcag catggcacgt
     1921 aatggagact accgcagttt ggtcattggg ctgcagtatg ctgggatccc cagtgttaac
     1981 tctttgcatt ctgtctacaa cttttgtgac aaaccctggg tgtttgccca gatggttcga
     2041 ctacacaaga agcttggaac agaggaattc cctctgattg atcagacttt ctatcccaat
     2101 cataaagaga tgctcagcag cacaacatac cctgtagttg tgaagatggg ccacgcacac
     2161 tctgggatgg gcaaggtcaa ggtagacaac caacatgact tccaggatat tgcaagtgtt
     2221 gtggcactga ctaagacata tgccactgct gagcccttca ttgatgctaa atacgatgtg
     2281 cgtgtccaga agattgggca gaactacaag gcctacatga ggacatcagt gtcagggaac
     2341 tggaagacca atacaggctc tgctatgctt gagcagattg ctatgtctga caggtacaag
     2401 ttgtgggtag acacgtgctc agagattttt gggggacttg acatctgcgc agtggaagca
     2461 ctgcatggca aggacggaag ggatcacatt attgaggtgg tgggctcctc catgccactc
     2521 attggggatc accaggatga agacaagcag ctcatcgtgg aacttgtggt caacaagatg
     2581 actcaggctc tgcctcggca gcgggatgct tcccctggca ggggttccca cagccagact
     2641 ccatccccag gagccctgcc cttgggccgc cagacctccc agcagcctgc aggacctcct
     2701 gctcaacaac gacccccacc ccagggaggc cctccacaac caggcccagg acctcagcgc
     2761 cagggacccc cgctgcaaca gcgcccaccc ccacaaggcc agcaacatct ttctggcctt
     2821 ggacccccag ctggcagccc tctgccccag cgcctaccaa gtcccacagc agcacctcag
     2881 cagtccgcct ctcaggccac accaatgacc cagggtcaag gccgccagtc acggccagtg
     2941 gcaggaggcc ccggagcacc tccagcagcc cgcccgccgg cctccccatc tccacagcgt
     3001 caggcagggc ccccacaggc tacccgtcag gcatctatct ctggtccagc tccaccgaag
     3061 gtctcaggag cctcacccgg agggcagcag cgccaaggcc ctccccagaa acccccaggc
     3121 cctgctggtc ccattcgtca ggccagccag gcaggtcccg gacctcgcac tgggccaccc
     3181 accacacagc agccccggcc cagcggccca ggtcctgctg gacgtcccac caaaccacag
     3241 ctggctcaga aacccagcca ggatgtgcca ccacccatca ttgctgctgc cgggggaccc
     3301 ccgcaccccc agctcaacaa atcccagtct ctgaccaatg ccttcaacct tccagagcca
     3361 gccccgccca ggcccagcct tagccaggat gaggtgaaag ctgagaccat ccgcagcctg
     3421 aggaagtctt tcgccagcct cttctccgac tgagtcgacg gtaccgcggg cccgggatcc
     3481 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3541 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3601 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3721 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3781 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3841 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3901 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3961 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4021 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4081 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4141 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     4201 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4261 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4321 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4561 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4621 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4681 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4741 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4801 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4861 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4921 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4981 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5041 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5101 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5161 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5221 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5281 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5341 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5401 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5461 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5521 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5581 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5641 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5701 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5761 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5821 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5881 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5941 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     6001 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6061 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6121 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6181 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6241 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6301 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6361 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6421 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6481 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6541 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6601 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6661 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6721 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6781 cctgattctg tggataaccg tattaccgcc atgcat
//
