LOCUS       sequence-172848-        6776 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 6776)
  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 6776)
  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: IDP431
FEATURES             Location/Qualifiers
     source          1..6776
                     /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"
     CDS             1459..1488
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1495..2040
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylcytosine 
                     derivatives"
                     /label=CLIP-tag(TM)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
                     VKEWLLAHEGHRLGKPGLG"
     regulatory      3157..3166
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3163..3228
                     /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    3564..3685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3601..3620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3655..3674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3692..4147)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3829..3850)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4041..4060
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4174..4278
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4280..4637
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4280..4471
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4303..4323)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4488..4623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4672..5466
                     /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             4672..5466
                     /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(4726..4745)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5336..5355
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5654..5673)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5698..5745
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6074..6662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6563..6582
                     /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 caccatgtac
     1321 cagaggatgc tccggtgcgg cgccgatctg ggatcgcccg ggggcggcag tggcggcggc
     1381 gcaggggggc gcctggccct gatctggata gtcccgctca ccctcggcgg cctcctagga
     1441 gtggcctggg gggaattcga acaaaaactc atctcagaag aggatctaca attcatggac
     1501 aaagactgcg aaatgaagcg caccaccctg gatagccctc tgggcaagct ggaactgtct
     1561 gggtgcgaac agggcctgca ccgtatcatc ttcctgggca aaggaacatc tgccgccgac
     1621 gccgtggaag tgcctgcccc agccgccgtg ctgggcggac cagagccact gatccaggcc
     1681 accgcctggc tcaacgccta ctttcaccag cctgaggcca tcgaggagtt ccctgtgcca
     1741 gccctgcacc acccagtgtt ccagcaggag agctttaccc gccaggtgct gtggaaactg
     1801 ctgaaagtgg tgaagttcgg agaggtcatc agcgagagcc acctggccgc cctggtgggc
     1861 aatcccgccg ccaccgccgc cgtgaacacc gccctggacg gaaatcccgt gcccattctg
     1921 atcccctgcc accgggtggt gcagggcgac agcgacgtgg ggccctacct gggcgggctc
     1981 gccgtgaaag agtggctgct ggcccacgag ggccacagac tgggcaagcc tgggctgggt
     2041 gtcgacggta ccgcatccag tttgggagcg caccacatcc accatttcca tggcagcagc
     2101 aagcatcatt cagtgcctat tgcaatctac aggtcaccag catccttgcg aggcggacac
     2161 gctgggacga cgtatatctt tagcaaaggt ggtggacaaa ttacatataa gtggcctcct
     2221 aatgaccgac ccagtacacg agcagacagg ctggccatcg ggtttagcac tgttcagaag
     2281 gaagcagtgt tggtgcgtgt ggacagttcc tcaggcctgg gtgactacct tgagctgcac
     2341 atacaccaag gaaaaattgg agttaaattt aatgttggga cagatgacat cgccattgag
     2401 gagtccaatg caatcattaa tgatggaaaa taccatgtag tgcgtttcac gaggagtggt
     2461 ggcaatgcca cgttacaggt ggacagctgg ccagtcatcg aacgataccc tgcagggcgt
     2521 cagctcacaa tcttcaatag ccaagcaacc ataataattg gcgggaaaga gcagggccag
     2581 cccttccagg gccagctctc tgggctttac tacaatggct tgaaagttct gaatatggcg
     2641 gcagagaacg atgccaacat cgccatagtg ggaaatgtga ggctggtcgg tgaagtgcct
     2701 tcctctatga caactgagtc gacagccact gccatgcagt ccgagatgtc cacctcaatc
     2761 atggagacca ccacaaccct ggctaccagc acagctcgac gaggaaagcc ccccacaaag
     2821 gagcctatca gccagaccac ggatgatatc cttgtggcct cggcagagtg tcccagcgac
     2881 gatgaggaca ttgacccctg tgagccgagc tcaggtgggt tagccaaccc cacccgagtg
     2941 ggcggccgcg aaccataccc aggctcggca gaggtgatcc gggagtctag cagtaccact
     3001 ggcatggtgg tggggattgt cgcagcagca gctctgtgca tcctcatcct cctctatgcc
     3061 atgtacaagt acaggaaccg ggatgaaggg tcgagcgctc caccgctcga cggagccgga
     3121 gctggcgctg gagctggagc cggagctggc gggctagcca ccatggacgc acaaacacga
     3181 cgacgtgagc gtcgcgctga gaaacaagct caatggaaag ctgcaaacct cgagccaccg
     3241 ctcgacggag ccggagctgg cgctggagct ggagccggag ctggcggtct agccacccca
     3301 ccgagcgctt accacgtgga tgagagtcga aactacatca gtaactcagc acagtccaat
     3361 ggggctgtgg tcaaggagat gcaacccagc agtgctaaaa gcgccaacaa aaacaagaag
     3421 aacaaggata aggagtatta tgtctaggga tcctctagat cataatcagc cataccacat
     3481 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3541 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3601 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3661 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3721 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3781 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3841 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3901 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3961 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4021 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4081 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4141 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4201 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4261 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4441 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4501 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4561 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4621 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4681 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4741 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4801 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4861 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4921 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4981 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5041 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5101 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5161 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5221 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5281 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5341 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5401 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5461 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5521 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5581 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5641 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5701 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5761 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5821 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5881 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5941 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6001 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6061 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6121 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6181 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6241 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6301 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6361 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6421 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6481 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6541 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6601 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6661 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6721 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
