LOCUS       sequence-169737-        5647 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 5647)
  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 5647)
  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..5647
                     /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"
     regulatory      1545..1554
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1681..1699)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1949..1968
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2143..2166
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2435..2556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2472..2491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2526..2545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2563..3018)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2700..2721)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2912..2931
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3045..3149
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3151..3508
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3151..3342
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3174..3194)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3359..3494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3543..4337
                     /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             3543..4337
                     /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(3597..3616)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4207..4226
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4525..4544)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4569..4616
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4945..5533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5434..5453
                     /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 cttataacct ccgaaaatgc tggcaccgtt ctctaagatc ggaagagcgg ttcagcagga
     1501 atgccgagac cgatctcgta tgccgtcttc tgcttggcgg ccgcgccacc atggcctcct
     1561 ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
     1621 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1681 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1741 agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1801 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1861 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1921 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1981 aggcctccac cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga
     2041 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca
     2101 agaagcccgt gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc
     2161 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactcctaat
     2221 gtacagccct gaaaaagggc tcgagccctg aaaaagggca attgccctga aaaagggcgt
     2281 cgacgccctg aaaaagggcg gtaccgcggg cccggggccg cgactctaga tcataatcag
     2341 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2401 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2461 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2521 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2581 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2641 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2701 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2761 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2821 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2881 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2941 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3001 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3121 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3181 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3241 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3301 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3361 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3421 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3481 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3541 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3601 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3661 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3721 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3781 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3841 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3901 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3961 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4021 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4081 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4141 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4201 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4261 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4321 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4381 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4441 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4501 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4561 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4621 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4681 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4741 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4801 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4861 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4921 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5101 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5161 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5221 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5281 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5341 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5401 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5461 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5521 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5581 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5641 catgcat
//
