LOCUS       sequence-463547-        4724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector containing mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4724)
  AUTHORS   Sandilands E, Freckmann EC, Cumming EM, Roman-Fernandez A, McGarry 
            L, Anand J, Galbraith L, Mason S, Patel R, Nixon C, Cartwright J, 
            Leung HY, Blyth K, Bryant DM
  TITLE     The small GTPase ARF3 controls invasion modality and metastasis by 
            regulating N-cadherin levels.
  JOURNAL   J Cell Biol. 2023 Apr 3;222(4):e202206115. doi: 
            10.1083/jcb.202206115. Epub 2023 Feb 28.
  PUBMED    36880595
REFERENCE   2  (bases 1 to 4724)
  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: "J Cell 
            Biol."; date: "2023-04-3"; volume: "222(4):e202206115. doi"; pages: 
            " 10.1083/jcb.202206115. Epub 2023 Feb 28"
COMMENT     Sequence Label: pmNeon Green-N1
FEATURES             Location/Qualifiers
     source          1..4724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        100..204
                     /gene="bla"
                     /label=AmpR promoter
     promoter        206..563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        206..397
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(229..249)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      414..549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             598..1392
                     /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             598..1392
                     /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(652..671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1262..1281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1580..1599)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1624..1671
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2000..2588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2489..2508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2763..3066
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3067..3270
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3221..3241
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3293..3373
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      3375..3384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3381..4091
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3636..3645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4214..4335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4251..4270)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4305..4324
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(4342..4724,1..73))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4479..4500)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4691..4710
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
       61 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      181 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
      241 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
      301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
      361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
      421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
      481 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
      541 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
      601 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
      661 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
      721 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
      781 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
      841 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
      901 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
      961 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     1021 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     1081 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     1141 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     1201 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     1261 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     1321 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     1381 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     1441 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     1501 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     1561 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     1621 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     1681 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     1741 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     1801 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     1861 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1921 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2221 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2281 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2341 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2401 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2461 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2521 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2581 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2641 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     2701 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     2761 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     2821 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     2881 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     2941 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     3001 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     3061 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     3121 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     3181 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     3241 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
     3301 taccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc
     3361 gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg gcctctctcc
     3421 cagcgacaca tgagttacac atctttggct ccatcaacgg tgtggacttt gacatggtgg
     3481 gtcagggcac cggcaatcca aatgatggtt atgaggagtt aaacctgaag tccaccaagg
     3541 gtgacctcca gttctccccc tggattctgg tccctcatat cgggtatggc ttccatcagt
     3601 acctgcccta ccctgacggg atgtcgcctt tccaggccgc catggtagat ggctccggct
     3661 accaagtcca tcgcacaatg cagtttgaag atggtgcctc ccttactgtt aactaccgct
     3721 acacctacga gggaagccac atcaaaggag aggcccaggt gaaggggact ggtttccctg
     3781 ctgacggtcc tgtgatgacc aactcgctga ccgctgcgga ctggtgcagg tcgaagaaga
     3841 cttaccccaa cgacaaaacc atcatcagta cctttaagtg gagttacacc actggaaatg
     3901 gcaagcgcta ccggagcact gcgcggacca cctacacctt tgccaagcca atggcggcta
     3961 actatctgaa gaaccagccg atgtacgtgt tccgtaagac ggagctcaag cactccaaga
     4021 ccgagctcaa cttcaaggag tggcaaaagg cctttaccga tgtgatgggc atggacgagc
     4081 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     4141 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4201 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4261 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4321 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4381 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4441 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4501 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4561 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4621 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4681 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcg
//
