LOCUS       sequence-172850-        7978 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 7978)
  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 7978)
  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: IDP432
FEATURES             Location/Qualifiers
     source          1..7978
                     /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      1309..1318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1456..1482
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1489..2034
                     /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"
     regulatory      2496..2505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4291..4300
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4297..4362
                     /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    4766..4887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4803..4822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4857..4876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4894..5349)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5031..5052)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5243..5262
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5376..5480
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5482..5839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5482..5673
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5505..5525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5690..5825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5874..6668
                     /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             5874..6668
                     /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(5928..5947)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6538..6557
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6856..6875)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6900..6947
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7276..7864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7765..7784
                     /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 ccgctagcgc caccatggca
     1321 cttcccagat gcatgtggcc aaattatgtt tggagagcta tgatggcatg tgtggtccac
     1381 aggggatccg gtgccccatt gactctctgc ttgttgggat gtttgctaca gacttttcac
     1441 gtactctctc aaaagtatcc atacgacgtt ccggactacg cagaattcat ggacaaagac
     1501 tgcgaaatga agcgcaccac cctggatagc cctctgggca agctggaact gtctgggtgc
     1561 gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa catctgccgc cgacgccgtg
     1621 gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc cactgatgca ggccaccgcc
     1681 tggctcaacg cctactttca ccagcctgag gccatcgagg agttccctgt gccagccctg
     1741 caccacccag tgttccagca ggagagcttt acccgccagg tgctgtggaa actgctgaaa
     1801 gtggtgaagt tcggagaggt catcagctac agccacctgg ccgccctggc cggcaatccc
     1861 gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc ccgtgcccat tctgatcccc
     1921 tgccaccggg tggtgcaggg cgacctggac gtggggggct acgagggcgg gctcgccgtg
     1981 aaagagtggc tgctggccca cgagggccac agactgggca agcctgggct gggtgtcgac
     2041 ttggatgatg tagacccatt ggttactact aactttggca agattagggg aattaagaaa
     2101 gaactcaata atgaaatttt gggtcctgtc attcagtttc ttggggttcc atatgccgct
     2161 ccaccaacag gagaacatcg tttccagcct ccagaaccac catctccctg gtctgacatc
     2221 cggaacgcca ctcagtttgc tcctgtatgt ccccagaata tcattgatgg cagattgcct
     2281 gaagttatgc ttcctgtgtg gttcactaat aacttggatg tggtttcatc atacgtccaa
     2341 gaccagagtg aagactgtct atacttaaac atctatgtcc caactgaaga tggtcccctt
     2401 acaaagaaac acacagatga tttaggtgat aatgacggtg ctgaagatga agatattcgg
     2461 gacagtgggg gtcccaaacc agtgatggtg tacatccatg gcggctctta catggaaggt
     2521 actggaaatc tgtatgatgg gagtgtcttg gcaagctatg gcaatgtgat cgtcatcaca
     2581 gtcaactatc ggcttggggt acttggcttc ttgagcacag gggatcaggc tgccaaagga
     2641 aactacgggc tccttgacct catccaggcc ctaagatgga ccagcgagaa cattgggttc
     2701 tttggtggtg accccttgcg aatcactgtg tttggatcag gcgctggggg ttcatgtgtc
     2761 aacctgctga ctttatccca ttattctgaa ggtaaccgtt ggagcaattc aaccaaagga
     2821 ctttttcaac gagcaatagc tcagagtgga acagcccttt ccagctgggc tgttagtttc
     2881 cagcctgcaa aatacgctag aattctggcc acaaaagttg gctgcaatgt ttcagataca
     2941 gtagagttag tagaatgcct gcagaagaag ccttacaaag aacttgttga tcaagatgtt
     3001 caaccagccc gataccacat agcctttgga cctgtgatcg atggtgatgt aataccagat
     3061 gaccctcaga tactgatgga acaaggagag ttcctcaact atgatataat gttgggagtt
     3121 aaccaagggg aagggttgaa gtttgtcgaa aatatagtag atagtgatga tggtgtatca
     3181 gccagtgatt tcgactttgc tgtttctaat tttgttgata atttatatgg atatcctgaa
     3241 ggcaaagatg ttttgagaga aaccattaaa ttcatgtata ctgactgggc tgatcgccat
     3301 aatcctgaaa ctagaaggaa gacattgttg gctttgttta cggaccatca atgggtagca
     3361 cctgctgtgg ccacagcaga ccttcactcg aactttggct cacctacata cttctatgcc
     3421 ttttatcatc attgccaaac agatcaagtt ccagcttggg ctgatgcagc tcatggggat
     3481 gaggttccct atgtgttggg aatccccatg attggcccta cagagttatt tccttgcaat
     3541 ttctccaaga atgatgtgat gttgagtgca gtagtaatga catactggac gaattttgct
     3601 aaaactggtg acccaaatca accagttcct caagacacaa aattcatcca taccaaaccc
     3661 aaccgctttg aagaagtagc atggaccaga tattcccaga aagaccagct ttatctccat
     3721 attggattaa aaccgagagt taaagagcat tacagagcca ataaggtaaa tctctggctg
     3781 gagctggtac ctcatctgca taatctcaat gacatttctc agtatacctc gacaacaact
     3841 aaagtgccat ccacggacat cactctcaga cctacaagga agaattccac accagtcaca
     3901 tcagcctttc ccactgccaa acaggatgat cccaagcaac aaccaagccc cttctcggtg
     3961 gatcagaggg actactccac agagctaagt gtcactatcg cagtgggggc ctctctgctg
     4021 tttctcaaca tcttggcttt tgcagccctg tactacaaga aggataagag gagacatgat
     4081 gtccaccgga ggtgcagccc tcagcgcacg accaccaacg acctaaccca tgctccagaa
     4141 gaggaaatta tgtctctcca aatgaagcac actgacttgg atcacgagtg tgagtccatc
     4201 catccacatg aggtggttct tcggaccagc gctccaccgc tcgacggagc cggagctggc
     4261 gctggagctg gagccggagc tggcgggcta gccaccatgg acgcacaaac acgacgacgt
     4321 gagcgtcgcg ctgagaaaca agctcaatgg aaagctgcaa acctcgagcc accgctcgac
     4381 ggagccggag ctggcgctgg agctggagcc ggagctggcg gtctagccac cccaccgagc
     4441 gctgcctgtc ccccagatta tactctagct atgaggaggt cacctgatga tattccacta
     4501 atgacaccta acaccatcac aatgattccc aacactatac cagggattca gcccttacat
     4561 acattcaaca catttactgg aggacagaat aatacactgc cccatcccca cccacacccc
     4621 cattcacatt caacaaccag ggtataggcg gccgctctag atcataatca gccataccac
     4681 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4741 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4801 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4861 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     4921 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4981 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5041 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5101 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5161 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5221 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5281 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     5341 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5401 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5461 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     5521 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5581 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5641 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5701 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5761 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5821 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     5881 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5941 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6001 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     6061 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6121 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6181 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6241 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6301 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6361 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     6421 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6481 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6541 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6601 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6661 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6721 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6781 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6841 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     6901 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     6961 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     7021 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     7081 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     7141 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7201 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7261 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7321 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7381 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7441 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7501 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7561 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7621 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7681 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7741 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7801 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7861 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7921 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
