LOCUS       sequence-127234-        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 505, Emission: 515. 
            This plasmid encodes dClover2 H231L based on Bajar et. al. Sci Rep. 
            2016..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4732)
  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: 
            "unpublished"
COMMENT     Sequence Label: Clover2-N1
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..25
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    192..313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(229..248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     283..302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(320..775)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(457..478)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     669..688
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        802..906
                     /gene="bla"
                     /label=AmpR promoter
     promoter        908..1265
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        908..1099
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(931..951)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1116..1251
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1300..2094
                     /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             1300..2094
                     /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(1354..1373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1964..1983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2282..2301)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2326..2373
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2702..3290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3191..3210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3465..3768
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3769..3972
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3923..3943
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3995..4075
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      4077..4086
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4083..4732,1..70)
                     /codon_start=1
                     /product="monomeric derivative of the bright green-yellow 
                     fluorescent protein Clover"
                     /label=mClover2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNSIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4128..4149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
       61 gtacaagtaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
      121 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
      181 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
      241 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
      301 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
      361 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
      421 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
      481 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
      541 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
      601 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
      661 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
      721 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
      781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      901 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
      961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1081 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     1141 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1201 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1261 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     1321 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     1381 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     1441 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     1501 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     1561 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     1621 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     1681 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     1741 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     1801 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     1861 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     1921 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     1981 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     2041 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     2101 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     2161 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     2221 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     2281 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     2341 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     2401 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     2461 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     2521 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     2581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3361 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     3421 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3481 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3541 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3601 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3661 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3721 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     3781 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     3841 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     3901 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     3961 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg
     4021 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     4081 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4141 acggcgacgt aaacggccac aagttcagcg tccgcggcga gggcgagggc gatgccacca
     4201 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4261 ccctcgtgac caccttcggc tacggcgtgg cctgcttcag ccgctacccc gaccacatga
     4321 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4381 ctttcaagga cgacggtacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4441 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4501 acaagctgga gtacaacttc aacagccact acgtctatat cacggccgac aagcagaaga
     4561 acagcatcaa ggctaacttc aagatccgcc acaacgttga ggacggcagc gtgcagctcg
     4621 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4681 actacctgag ccatcagtcc gccctgagca aagaccccaa cgagaagcgc ga
//
