LOCUS       sequence-172860-        5590 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 5590)
  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 5590)
  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: IDP379
FEATURES             Location/Qualifiers
     source          1..5590
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      21..30
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(157..175)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     425..444
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     619..642
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    911..1032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(948..967)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1002..1021
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1039..1494)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1176..1197)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1388..1407
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1521..1625
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1627..1984
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1627..1818
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1650..1670)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1835..1970
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2019..2813
                     /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             2019..2813
                     /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(2073..2092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2683..2702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3001..3020)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3045..3092
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3421..4009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3910..3929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4223..5401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4454..5392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5349..5369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gtcttctgct tggcggccgc gccaccatgg cctcctccga ggacgtcatc aaggagttca
       61 tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      121 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
      181 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt ccagtacggc tccaaggcct
      241 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      301 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      361 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
      421 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cggatgtacc
      481 ccgaggacgg cgccctgaag ggcgagatca agatgaggct gaagctgaag gacggcggcc
      541 actacgacgc cgaggtcaag accacctaca tggccaagaa gcccgtgcag ctgcccggcg
      601 cctacaagac cgacatcaag ctggacatca cctcccacaa cgaggactac accatcgtgg
      661 aacagtacga gcgcgccgag ggccgccact cctaatgtac agccctgaaa aagggctcga
      721 gccctgaaaa agggcaattg ccctgaaaaa gggcgtcgac gccctgaaaa agggcggtac
      781 cgcgggcccg gggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
      841 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
      901 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
      961 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1021 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     1081 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     1141 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     1201 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1261 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     1321 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     1381 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     1441 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     1501 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1561 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1621 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     1681 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     1741 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1801 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     1861 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     1921 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     1981 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2041 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2101 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2161 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     2221 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2281 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2341 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2401 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2461 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2521 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     2581 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2641 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2701 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     2761 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     2821 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     2881 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     2941 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     3001 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     3061 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3121 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3181 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3241 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     3301 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3361 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3421 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3481 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3541 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3601 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3661 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3721 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3781 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3841 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3901 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3961 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4021 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4081 gttatcccct gattctgtgg ataaccgtat taccgccatg ctcctgcagg acacgtaagc
     4141 tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
     4201 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     4261 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     4321 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     4381 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     4441 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     4501 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
     4561 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
     4621 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
     4681 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     4741 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     4801 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     4861 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     4921 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     4981 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     5041 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     5101 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     5161 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     5221 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     5281 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     5341 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     5401 aggtacgacc accgctagcg ctggtcggta cggtaacgga gagtccgtcc cctcttatcc
     5461 tcggcgttgt gtgtcaaatg gcgtagatct ggattgactc tatgacggta tctgctgatc
     5521 ggtagggaga ccgagaatct atcgggctaa cgcgtcttat aacctccgaa aatgctggca
     5581 ccgttctcta
//
