LOCUS       sequence-169735-        7325 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 7325)
  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 7325)
  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: IDP254
FEATURES             Location/Qualifiers
     source          1..7325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        112..1290
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          343..1281
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1238..1258
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2135..2161
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2168..2713
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             complement(2993..3004)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3242..3251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3248..3313
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     regulatory      3371..3380
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3377..3442
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     regulatory      3500..3509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3506..3571
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     regulatory      3629..3638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3635..3700
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     polyA_signal    4113..4234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4150..4169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4204..4223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4241..4696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4378..4399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4590..4609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4723..4827
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4829..5186
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4829..5020
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4852..4872)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5037..5172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5221..6015
                     /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             5221..6015
                     /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(5275..5294)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5885..5904
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6203..6222)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6247..6294
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6623..7211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7112..7131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tcctgcagga cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag
       61 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
      121 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
      181 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      241 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      301 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
      361 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
      421 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      481 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      541 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      601 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      661 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      721 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
      781 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
      841 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
      901 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
      961 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1021 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1081 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1141 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1201 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1261 agtttttttc ttccatttca ggtgtcgtga ggtacgacca ccgcctaggg ccaccatggc
     1321 acttcctaga tgtatgtggc ctaattatgt atggcgagct atgatggcct gcgtggtgca
     1381 ccgagggagt ggcgccccac ttacactctg cctgttgggt tgcctcctcc agactttcca
     1441 cgttctttca cagaagctgg atgacatgtc caagggagaa gagttgttta ctggcgtagt
     1501 ccctattctc gtggaactcg atggtgacgt gaatggccat aagttttctg tcagaggaga
     1561 gggagaaggc gatgccacca tcgggaaact cacgctgaaa ttcatctgta ccactggaaa
     1621 acttcccgtg ccttggccaa ccctcgtgac aacactcacc tacggggtgc aatgtttctc
     1681 tcggtacccg gaccatatga agaggcatga cttcttcaag agtgccatgc ccgagggtta
     1741 cgttcaggag cgcaccatct cttttaagga cgatggcaaa tataagacaa gagcagtcgt
     1801 caagttcgag ggtgatacac tcgttaaccg catcgagctc aaaggcaccg attttaagga
     1861 ggacggaaat atcctgggac acaaattgga gtacaacttc aacagtcaca acgtgtatat
     1921 tacagcagat aagcagaaga atggcataaa ggccaatttc acggtaagac ataatgtcga
     1981 ggatggcagt gtccagctgg cagaccacta ccagcaaaac acgcccattg gcgatggacc
     2041 tgttctcctg ccagacaacc actacctcag tacccaaaca gtcctgtcca aggaccctaa
     2101 tgagaaaggg acgggcggtg gcggatctct cgagtatcca tacgacgttc cggactacgc
     2161 agaattcatg gacaaagact gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa
     2221 gctggaactg tctgggtgcg aacagggcct gcaccgtatc atcttcctgg gcaaaggaac
     2281 atctgccgcc gacgccgtgg aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc
     2341 actgatgcag gccaccgcct ggctcaacgc ctactttcac cagcctgagg ccatcgagga
     2401 gttccctgtg ccagccctgc accacccagt gttccagcag gagagcttta cccgccaggt
     2461 gctgtggaaa ctgctgaaag tggtgaagtt cggagaggtc atcagctaca gccacctggc
     2521 cgccctggcc ggcaatcccg ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc
     2581 cgtgcccatt ctgatcccct gccaccgggt ggtgcagggc gacctggacg tggggggcta
     2641 cgagggcggg ctcgccgtga aagagtggct gctggcccac gagggccaca gactgggcaa
     2701 gcctgggctg ggtggcggcg gtggtagcct cgagctggtg ccccacttgc ataacctcaa
     2761 tgacatcagc caatacacaa gtacaaccac caaggtccca agcactgaca tcacccttcg
     2821 accaactcgg aaaaactcaa cccctgtgac cagcgcattt ccaaccgcta agcaagacga
     2881 ccccaaacaa cagcctagtc catttagcgt agaccaaagg gactacagca cagagctgtc
     2941 cgtgaccata gctgtaggag cttccctgct tttcctgaac attctggctt tcgcggccct
     3001 ctattacaag aaggacaaaa ggcgccacga cgtacatagg agatgtagtc cgcaacgaac
     3061 tacaaccaat gatcttacgc acgcacctga ggaggagatc atgagtcttc agatgaaaca
     3121 cacagacctg gaccacgagt gtgaatccat ccatccccat gaggtggtgc tgaggacaag
     3181 cgctccaccg ctcgacggag ccggagctgg cgctggagct ggagccggag ctggcgggct
     3241 agccaccatg gacgcacaaa cacgacgacg tgagcgtcgc gctgagaaac aagctcaatg
     3301 gaaagctgca aacccaccgc tcgacggagc cggagctggc gctggagctg gagccggagc
     3361 tggcggtcta gccaccatgg acgcacaaac acgacgacgt gagcgtcgcg ctgagaaaca
     3421 agctcaatgg aaagctgcaa acccaccgct cgacggagcc ggagctggcg ctggagctgg
     3481 agccggagct ggcggtctag ccaccatgga cgcacaaaca cgacgacgtg agcgtcgcgc
     3541 tgagaaacaa gctcaatgga aagctgcaaa cccaccgctc gacggagccg gagctggcgc
     3601 tggagctgga gccggagctg gcggtctagc caccatggac gcacaaacac gacgacgtga
     3661 gcgtcgcgct gagaaacaag ctcaatggaa agctgcaaac ccaccgctcg agccaccgct
     3721 cgacggagcc ggagctggcg ctggagctgg agccggagct ggcggtctag ccaccccacc
     3781 gagcgctgca tgccctccgg actataccct ggcaatgcgc cgaagcccag acgacattcc
     3841 cctgatgaca cccaatacca taaccatgat tcccaacact attccaggaa tccagcctct
     3901 ccacactttc aacacattca ctggtggaca gaataatact ctccctcatc cgcatccaca
     3961 cccccactcc cacagcacaa cacgggtgta ggcggccgcg actctagatc ataatcagcc
     4021 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4081 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4141 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4201 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     4261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4681 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4801 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4861 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4921 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4981 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5041 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5101 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5161 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     5221 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5281 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5341 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5401 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5461 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5521 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5581 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5641 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5701 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     5761 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5821 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5881 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5941 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6001 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6061 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6121 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6181 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6241 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6301 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6361 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     6421 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     6481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     6781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7261 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     7321 tgcat
//
