LOCUS       sequence-389150-        4961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane localizing mNeonGreen using Human LCK proto-oncogene
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4961)
  TITLE     Fluorescent Membrane and Nuclear Localization Proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4961)
  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            "Unpublished"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: Membrane mNeonGreen
FEATURES             Location/Qualifiers
     source          1..4961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        191..209
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     191..208
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      330..339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      393..402
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             399..1109
                     /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      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1169..1303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1193..1212)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1247..1266
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1416..1436)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1416..1434)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1455..1471)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1455..1471)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1468..1490)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1479..1495
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1503..1509,1510..1527,1528..1533))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1548..1569
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1686..1703)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1857..2445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1937..1956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2616..3407,3408..3476))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3239..3258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3477..3581)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3607..4062)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3744..3765)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3956..3975
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4133..4961,1..156)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg acgtcaatgg
       61 ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa
      121 tgggaggtct atataagcaa tgctcgttta gggaaccgcc attctgcctg gggacgtcgg
      181 agcaagcttg atttaggtga cactatagaa tacaagctac ttgttctttt tgcaggatcc
      241 actagtggcg cgccattaat taaaggccgg ccagcgatcg catgggctgc atcaagagca
      301 agcgcaagga caacctgaac gacgacgagg ccgccatggg ttgtattaaa tcaaaacgaa
      361 aagataattt gaatgatgat gaagcaccgg tcgccaccat ggtgagcaag ggcgaggagg
      421 ataacatggc ctctctccca gcgacacatg agttacacat ctttggctcc atcaacggtg
      481 tggactttga catggtgggt cagggcaccg gcaatccaaa tgatggttat gaggagttaa
      541 acctgaagtc caccaagggt gacctccagt tctccccctg gattctggtc cctcatatcg
      601 ggtatggctt ccatcagtac ctgccctacc ctgacgggat gtcgcctttc caggccgcca
      661 tggtagatgg ctccggctac caagtccatc gcacaatgca gtttgaagat ggtgcctccc
      721 ttactgttaa ctaccgctac acctacgagg gaagccacat caaaggagag gcccaggtga
      781 aggggactgg tttccctgct gacggtcctg tgatgaccaa ctcgctgacc gctgcggact
      841 ggtgcaggtc gaagaagact taccccaacg acaaaaccat catcagtacc tttaagtgga
      901 gttacaccac tggaaatggc aagcgctacc ggagcactgc gcggaccacc tacacctttg
      961 ccaagccaat ggcggctaac tatctgaaga accagccgat gtacgtgttc cgtaagacgg
     1021 agctcaagca ctccaagacc gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg
     1081 tgatgggcat ggacgagctg tacaagtaac atcgattgga attcaaggcc tctcgagcct
     1141 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     1201 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1261 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1321 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     1381 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1441 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1501 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1561 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1621 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1681 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1741 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1801 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1861 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1921 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1981 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2041 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2101 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2161 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2221 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2281 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2341 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2401 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2461 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2521 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2581 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2641 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2701 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2761 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2821 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2881 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2941 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3001 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3061 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3121 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3181 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3241 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3301 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3361 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3421 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3481 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3541 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3601 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3661 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3721 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3781 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3841 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3901 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3961 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4021 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     4081 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
     4141 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     4201 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     4261 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
     4321 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     4381 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     4441 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     4501 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     4561 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     4621 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     4681 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     4741 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     4801 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     4861 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     4921 attggaagta cgccagggta cattggcagt actcccattg a
//
