LOCUS       sequence-172854-        5677 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 5677)
  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 5677)
  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: IDP378
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      17..26
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(23..25,26..28,29..742)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(68..89)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    958..1079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(995..1014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1049..1068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1086..1541)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1223..1244)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1435..1454
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1568..1672
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1674..2031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1674..1865
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1697..1717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1882..2017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2066..2860
                     /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             2066..2860
                     /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(2120..2139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2730..2749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3048..3067)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3092..3139
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3468..4056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3957..3976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4270..5448
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4501..5439
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5396..5416
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ccgtatcatt acgcgtgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
       61 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgaggggcga
      121 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      181 gctgcccgtg ccctggccca ccctcgtgac caccctgagc cacggcgtgc agtgcttcgc
      241 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      301 cgtccaggag cgcaccatct tcttcaagga cgacggcacc tacaagaccc gcgccgaggt
      361 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcgtcg acttcaagga
      421 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acatctatat
      481 catggccgtc aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacgtgga
      541 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      601 cgtgctgctg cccgacagcc actacctgag cacccagtcc gtgctgagca aagaccccaa
      661 cgagaagcgc gatcacatgg tcctgctgga gttccgcacc gccgccggga tcactctcgg
      721 catggacgag ctgtataagt aatgtacagc cctgaaaaag ggctcgagcc ctgaaaaagg
      781 gcaattgccc tgaaaaaggg cgtcgacgcc ctgaaaaagg gcggtaccgc gggcccgggg
      841 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
      901 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
      961 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1021 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     1081 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     1141 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     1201 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     1261 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     1321 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     1381 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     1441 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1501 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     1561 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1621 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     1681 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     1741 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     1801 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     1861 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     1921 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     1981 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     2041 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     2101 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     2161 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     2221 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     2281 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     2341 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     2401 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     2461 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     2521 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     2581 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     2641 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     2701 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     2761 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     2821 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     2881 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     2941 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     3001 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     3061 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     3121 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     3181 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     3241 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     3301 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3361 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3421 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3481 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3541 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3601 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3661 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3721 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3781 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3841 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3901 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3961 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4021 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4081 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4141 tctgtggata accgtattac cgccatgctc ctgcaggaca cgtaagcttt gcaaagatgg
     4201 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
     4261 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     4321 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
     4381 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     4441 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     4501 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
     4561 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
     4621 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
     4681 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
     4741 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
     4801 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
     4861 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
     4921 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     4981 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     5041 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     5101 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     5161 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     5221 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     5281 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     5341 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     5401 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg tacgaccacc
     5461 gctagccgga cgatgccgtc ctcgtagttc gggcatgtac tggagccgag aggtaacctc
     5521 attataatcg ttcgctattc agggattgac caacaccgga aaacatctca cttgaagtaa
     5581 tgtatacgac agagtcgcgg ccgccagtgc ttgggcaacc ggtggacacg tcttatgtga
     5641 gatcggaaga gcgtcgtgta gggaaagagt gtagatc
//
