LOCUS       sequence-267741-        4746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Peroxisomes, Excitation: 485, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4746)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4746)
  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            "unpublished"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: sfGFP-Peroxisomes-2
FEATURES             Location/Qualifiers
     source          1..4746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        149..452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        453..656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     607..627
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    679..759
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      761..770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(767..769,770..772,773..1483)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(812..833)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1420..1441
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1622..1743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1659..1678)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1713..1732
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1750..2205)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1887..1908)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2099..2118
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2232..2336
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2338..2695
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2338..2529
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2361..2381)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2546..2681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2730..3524
                     /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             2730..3524
                     /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(2784..2803)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3394..3413
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3712..3731)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3756..3803
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4132..4720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4621..4640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
       61 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
      121 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      181 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      241 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      301 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      361 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      421 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      481 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      541 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      601 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      661 tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat
      721 tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg
      781 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      841 ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc accaacggca agctgaccct
      901 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      961 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcgcc acgacttctt
     1021 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcagcttca aggacgacgg
     1081 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1141 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1201 cttcaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     1261 cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca
     1321 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1381 gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1441 gaccgccgcc gggatcactc acggcatgga cgagctgtac aagtccggat ccaagctgta
     1501 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     1561 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1621 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1681 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1741 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1801 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1861 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1921 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1981 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2041 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     2101 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     2161 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     2221 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2281 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     2341 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2401 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2461 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2521 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2581 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2641 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     2701 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     2761 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     2821 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     2881 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     2941 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3001 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3061 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3121 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3181 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3241 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3301 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3361 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3421 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3481 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3541 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     3601 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3661 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     3721 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3781 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3841 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3901 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3961 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     4021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acacggcctt
     4741 tttacg
//
