LOCUS       sequence-169732-        5788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for cell culture expression of SYNseq components.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5788)
  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 5788)
  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..5788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        108..1286
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          339..1277
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1234..1254
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     gap             1463..1492
                     /estimated_length=30
     regulatory      1572..1581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1578..2291
                     /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(1723..1741)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1991..2010
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2576..2697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2613..2632)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2667..2686
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2704..3159)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2841..2862)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3053..3072
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3186..3290
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3292..3649
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3292..3483
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3315..3335)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3500..3635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3684..4478
                     /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             3684..4478
                     /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(3738..3757)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4348..4367
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4666..4685)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4710..4757
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5086..5674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5575..5594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tcctgcagga taagctttgc aaagatggat aaagttttaa acagagagga
       61 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
      121 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      181 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      241 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      301 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
      361 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
      421 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
      481 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
      541 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
      601 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
      661 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
      721 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
      781 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
      841 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
      901 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
      961 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     1021 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     1081 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     1141 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     1201 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     1261 tttttcttcc atttcaggtg tcgtgaggta cgaccaccgc tagcgctggt cggtacggta
     1321 acggagagtc cgtcccctct tatcctcggc gttgtgtgtc aaatggcgta gatctggatt
     1381 gactctatga cggtatctgc tgatcggtag ggagaccgag aatctatcgg gctaacgcgt
     1441 aacctccgaa aatgctggca ccnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnagatcgga
     1501 agagcggttc agcaggaatg ccgagaccga tctcgtatgc cgtcttctgc ttggcggccg
     1561 cactaatcta ggccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     1621 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1681 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1741 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     1801 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     1861 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     1921 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     1981 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     2041 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     2101 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2161 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     2221 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2281 acaagtctta gagatcttac ctaattgccg tcgtgaggta cgaccaccgc tagctgtaca
     2341 gccctgaaaa agggctcgag ccctgaaaaa gggcaattgc cctgaaaaag ggcgtcgacg
     2401 ccctgaaaaa gggcggaatt catgtccaat ttactgactg tacaccaaaa tttgcctgcc
     2461 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2521 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2581 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2641 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     2701 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2761 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2821 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2881 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2941 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3001 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3061 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3121 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3181 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3241 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3301 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3361 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3421 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3481 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3541 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3601 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     3661 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3721 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3781 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3841 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     3901 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3961 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4021 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4081 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4141 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4201 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4261 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4321 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4381 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4441 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4501 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4561 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4621 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     4681 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4741 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4801 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4861 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4921 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4981 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5041 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5101 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5161 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5221 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5281 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5341 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5401 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5461 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5521 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5581 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5641 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5701 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5761 tgtggataac cgtattaccg ccatgcat
//
