LOCUS       sequence-172852-        5754 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  plasmid for cell culture expression of SYNseq components.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5754)
  AUTHORS   Peikon ID, Kebschull JM, Vagin VV, Ravens DI, Sun YC, Brouzes E, 
            Correa IR Jr, Bressan D, Zador AM
  TITLE     Using high-throughput barcode sequencing to efficiently map 
            connectomes.
  JOURNAL   Nucleic Acids Res. 2017 Jul 7;45(12):e115. doi: 10.1093/nar/gkx292.
  PUBMED    28449067
REFERENCE   2  (bases 1 to 5754)
  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: "Nucleic 
            Acids Res."; date: "2017-07-7"; volume: "45(12):e115. doi"; pages: "
            10.1093/nar/gkx292"
COMMENT     Sequence Label: IDP456
FEATURES             Location/Qualifiers
     source          1..5754
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      31..40
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             37..750
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKS"
     primer_bind     complement(182..200)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     450..469
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1035..1156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1072..1091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1126..1145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1163..1618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1300..1321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1512..1531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1645..1749
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1751..2108
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1751..1942
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1774..1794)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1959..2094
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2143..2937
                     /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             2143..2937
                     /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(2197..2216)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2807..2826
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3125..3144)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3169..3216
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3545..4133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4034..4053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4347..5525
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4578..5516
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5473..5493
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gtcttctgct tggcggccgc actaatctag gccaccatgg tgagcaaggg cgaggaggat
       61 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
      121 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
      181 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
      241 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
      301 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
      361 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
      421 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
      481 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
      541 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
      601 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
      661 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
      721 ggcggcatgg acgagctgta caagtcttag agatcttacc taattgccgt cgtgaggtac
      781 gaccaccgct agctgtacag ccctgaaaaa gggctcgagc cctgaaaaag ggcaattgcc
      841 ctgaaaaagg gcgtcgacgc cctgaaaaag ggcggaattc atgtccaatt tactgactgt
      901 acaccaaaat ttgcctgccg cgactctaga tcataatcag ccataccaca tttgtagagg
      961 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     1021 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1081 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1141 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     1201 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1261 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1321 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1381 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1441 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1501 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1561 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     1621 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1681 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1741 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     1801 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1861 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1921 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1981 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2041 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2101 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2161 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2221 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2281 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     2341 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2401 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2461 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     2521 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     2581 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     2641 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     2701 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     2761 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     2821 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     2881 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     2941 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3001 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3061 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     3121 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3181 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     3241 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     3301 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     3361 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     3421 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3481 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3541 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3601 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3661 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4021 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4081 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4141 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4201 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgctcctg caggacacgt
     4261 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     4321 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     4381 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     4441 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4501 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4561 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     4621 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     4681 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     4741 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     4801 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     4861 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     4921 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     4981 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     5041 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     5101 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     5161 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     5221 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     5281 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     5341 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     5401 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     5461 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     5521 cgtgaggtac gaccaccgct agcgctggtc ggtacggtaa cggagagtcc gtcccctctt
     5581 atcctcggcg ttgtgtgtca aatggcgtag atctggattg actctatgac ggtatctgct
     5641 gatcggtagg gagaccgaga atctatcggg ctaacgcgta acctccgaaa atgctggcac
     5701 catatcgtga attgtattta aggatgacgt aagatcggaa gagcggttca gcag
//
