LOCUS       sequence-217928-        5393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient/stable overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5393)
  AUTHORS   Niewiadomski P, Proszynski TJ
  TITLE     LL5-beta interacts with uncharacterised protein 2410002F23Rik 
            (LL5BIP)
  JOURNAL   Matters v. 2, n. 11  (Nov 17, 2016)
REFERENCE   2  (bases 1 to 5393)
  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: "Matters v. 
            2, n. 11  (Nov 17, 2016)"
COMMENT     Sequence Label: pSYN-LL5BIP-GFP
FEATURES             Location/Qualifiers
     source          1..5393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1371..1373,1374..1376,1377..2090)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1371..1373,1374..1376,1377..2087)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1416..1437)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1677..1696)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2024..2045
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2224..2345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2261..2280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2315..2334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2352..2807)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2489..2510)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2701..2720
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2834..2938
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2940..3297
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2940..3131
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2963..2983)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3148..3283
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3332..4126
                     /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             3332..4126
                     /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(3386..3405)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3996..4015
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4314..4333)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4358..4405
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4734..5322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5223..5242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gaattcaagc tgctcgagct caagcttggt
      481 accgagctcg gatccacggc cggccaacca tggctgagag agtgctggtg cccacccaga
      541 taggccgggg ggaccgctac tacacataca cggagttgtt ggctatctca cggcgtttca
      601 agcagaaccc caatgagctc atggtcacct ggatcctgcg ggtgtatgac cagggaggcc
      661 ctgccctgtc cctgaattct ggggagctgg ggctgctggg tgacctcacc catgatgcca
      721 tctttaacta ccgctgcaag gccctgcggg gggctggctg ccagactctc ctgagctggc
      781 tgctgcaggc ctggcgtcag cgttgggaat cctccctgca ttttgaggcc accgagttac
      841 ccttcaggcc ctggaccacc atggaggaag gcatccagtt ggtgcgtgag ctgggcatga
      901 ttgagtggat ctaccttgac ccagaagggc ccgtggacct ggccccagag gatgtggcct
      961 tcactcaagg cctgcagcgg cgcctgctca cagcagcccc ctctgagctg cggctttcac
     1021 tggtcagcct gctggtacgt ggcatgacag tgttggaggc tgtgatggag atccagacta
     1081 ttgctgacgt gggtctgctc tggcgccaga gccatccagg ccgcaccaag ctcatgttgg
     1141 ggcccaaccc aactcgtaag gacctcctag gctggctgct cagccacggt gtaccccggg
     1201 agcaagtaga cagacagccc accaaggtac tcctagaact ctacatcaaa gaagccaagc
     1261 gcagtcgcgg ccaccccaac tatgggctga acgaggagca gcccccacca cccccttact
     1321 ccgaccaggc ctgtggggaa gagcaaccgg tgcgtcatga cggcgcgcct atggtgagca
     1381 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1441 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1501 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1561 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1621 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1681 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1741 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1801 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1861 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1921 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1981 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2041 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa tctaggatcc
     2101 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2161 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2221 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2281 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2341 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2401 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2461 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2521 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2581 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2641 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2701 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2761 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2821 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2881 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     2941 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3001 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3061 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3121 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3181 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3241 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3301 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3361 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3421 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3481 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3541 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3601 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3661 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3721 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3781 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3841 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3901 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3961 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4021 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4081 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4141 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4201 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4261 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4321 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4381 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4441 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4501 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4561 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4621 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4681 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4741 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4801 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4861 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4921 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4981 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5041 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5101 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5161 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5221 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5281 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5341 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcgt atg
//
