LOCUS       sequence-389155-        4970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane localizing EGFP using Human LCK proto-oncogene 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4970)
  TITLE     Fluorescent Membrane and Nuclear Localization Proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4970)
  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            "Unpublished"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 EGFP
FEATURES             Location/Qualifiers
     source          1..4970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..55
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     37..54
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      176..185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      239..248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(245..247,248..250,251..964)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(290..311)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(551..570)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     898..919
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1024..1158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1048..1067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1102..1121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1271..1291)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1271..1289)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1310..1326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1310..1326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1323..1345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1334..1350
                     /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(1358..1364,1365..1382,1383..1388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1403..1424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1541..1558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1712..2300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1792..1811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2471..3262,3263..3331))
                     /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     3094..3113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3332..3436)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3462..3917)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3599..3620)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3811..3830
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(3988..4970,1..2)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca
       61 agctacttgt tctttttgca ggatccacta gtggcgcgcc attaattaaa ggccggccag
      121 cgatcgcatg ggctgcatca agagcaagcg caaggacaac ctgaacgacg acgaggccgc
      181 catgggttgt attaaatcaa aacgaaaaga taatttgaat gatgatgaag caccggtcgc
      241 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      301 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      361 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      421 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      481 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      541 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      601 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
      661 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
      721 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
      781 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
      841 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
      901 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
      961 gtaacatcga ttggaattca aggcctctcg agcctctaga actatagtga gtcgtattac
     1021 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1081 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1141 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1201 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
     1261 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     1321 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa
     1381 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1441 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1501 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1561 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1621 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1681 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1741 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1801 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1861 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1921 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1981 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2041 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2101 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2161 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2221 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2281 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2341 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2401 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2461 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2521 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2581 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2641 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2701 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2761 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2821 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2881 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2941 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3001 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3061 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3121 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3181 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3241 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3301 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3361 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3421 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     3481 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3541 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3601 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3661 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3721 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3781 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3841 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3901 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3961 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac
     4021 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
     4081 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
     4141 ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc
     4201 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
     4261 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
     4321 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
     4381 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
     4441 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
     4501 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
     4561 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
     4621 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
     4681 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
     4741 atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg
     4801 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
     4861 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     4921 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga
//
