LOCUS       sequence-140287-        6294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tight Junctions, Excitation: 487, Emission: 509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  TITLE     Davidson mEmerald
REFERENCE   2  (bases 1 to 6294)
  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"
COMMENT     Sequence Label: mEmerald-Occludin-C-14
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(620..622,623..625,626..1336)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(665..686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(926..945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1273..1294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             1402..1410
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3089..3210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3126..3145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3180..3199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3217..3672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3354..3375)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3566..3585
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3699..3803
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3805..4162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3805..3996
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3828..3848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4013..4148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4197..4991
                     /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             4197..4991
                     /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(4251..4270)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4861..4880
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5179..5198)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5223..5270
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5599..6187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6088..6107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      781 gcccgtgccc tggcccaccc tcgtgaccac cttgacctac ggcgtgcagt gcttcgcccg
      841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaagg tctatatcac
     1081 cgccgacaag cagaagaacg gcatcaaggt gaacttcaag acccgccaca acatcgagga
     1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1201 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1321 ggacgagctg tacaagtccg gactcagatc tcgaggaagc ggcggatctg ggtccggatc
     1381 atccaggcct cttgaaagtc cacctcctta caggcctgat gaattcaaac cgaatcatta
     1441 tgcaccaagc aatgacatat atggtggaga gatgcatgtt cgaccaatgc tctctcagcc
     1501 agcctactct ttttacccag aagatgaaat tcttcacttc tacaaatgga cctctcctcc
     1561 aggagtgatt cggatcctgt ctatgctcat tattgtgatg tgcattgcca tctttgcctg
     1621 tgtggcctcc acgcttgcct gggacagagg ctatggaact tcccttttag gaggtagtgt
     1681 aggctaccct tatggaggaa gtggctttgg tagctacgga agtggctatg gctatggcta
     1741 tggttatggc tatggctacg gaggctatac agacccaaga gcagcaaagg gcttcatgtt
     1801 ggccatggct gccttttgtt tcattgccgc gttggtgatc tttgttacca gtgttataag
     1861 atctgaaatg tccagaacaa gaagatacta cttaagtgtg ataatagtga gtgctatcct
     1921 gggcatcatg gtgtttattg ccacaattgt ctatataatg ggagtgaacc caactgctca
     1981 gtcttctgga tctctatatg gttcacaaat atatgccctc tgcaaccaat tttatacacc
     2041 tgcagctact ggactctacg tggatcagta tttgtatcac tactgtgttg tggatcccca
     2101 ggaggccatt gccattgtac tggggttcat gattattgtg gcttttgctt taataatttt
     2161 ctttgctgtg aaaactcgaa gaaagatgga caggtatgac aagtccaata ttttgtggga
     2221 caaggaacac atttatgatg agcagccccc caatgtcgag gagtgggtta aaaatgtgtc
     2281 tgcaggcaca caggacgtgc cttcaccccc atctgactat gtggaaagag ttgacagtcc
     2341 catggcatac tcttccaatg gcaaagtgaa tgacaagcgg ttttatccag agtcttccta
     2401 taaatccacg ccggttcctg aagtggttca ggagcttcca ttaacttcgc ctgtggatga
     2461 cttcaggcag cctcgttaca gcagcggtgg taactttgag acaccttcaa aaagagcacc
     2521 tgcaaaggga agagcaggaa ggtcaaagag aacagagcaa gatcactatg agacagacta
     2581 cacaactggc ggcgagtcct gtgatgagct ggaggaggac tggatcaggg aatatccacc
     2641 tatcacttca gatcaacaaa gacaactgta caagaggaat tttgacactg gcctacagga
     2701 atacaagagc ttacaatcag aacttgatga gatcaataaa gaactctccc gtttggataa
     2761 agaattggat gactatagag aagaaagtga agagtacatg gctgctgctg atgaatacaa
     2821 tagactgaag caagtgaagg gatctgcaga ttacaaaagt aagaagaatc attgcaagca
     2881 gttaaagagc aaattgtcac acatcaagaa gatggttgga gactatgata gacagaaaac
     2941 atagggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     3001 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3061 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3121 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3181 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     3241 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3301 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3361 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3421 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3481 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3541 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3601 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3661 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3721 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3781 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3841 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4081 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4141 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4201 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4261 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4321 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4381 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4441 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4501 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4561 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4621 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4681 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4741 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4801 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4861 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4921 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4981 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5041 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5101 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5161 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5221 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5281 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5341 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5401 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5461 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5521 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5581 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5641 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5701 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5761 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5821 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5881 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5941 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6001 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6061 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6121 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6181 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6241 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgc
//
