LOCUS       sequence-257392-        6244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tight Junctions, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6244)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 6244)
  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: mCherry-Occludin-N-10
FEATURES             Location/Qualifiers
     source          1..6244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     RBS             644..652
                     /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)"
     regulatory      2208..2217
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2214..2924
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2359..2377)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2627..2646
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3047..3168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3084..3103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3138..3157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3175..3630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3312..3333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3524..3543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3657..3761
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3763..4120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3763..3954
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3786..3806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3971..4106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4155..4949
                     /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             4155..4949
                     /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(4209..4228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4819..4838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5137..5156)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5181..5228
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5557..6145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6046..6065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgccaccatg tcatccaggc ctcttgaaag tccacctcct tacaggcctg
      661 atgaattcaa accgaatcat tatgcaccaa gcaatgacat atatggtgga gagatgcatg
      721 ttcgaccaat gctctctcag ccagcctact ctttttaccc agaagatgaa attcttcact
      781 tctacaaatg gacctctcct ccaggagtga ttcggatcct gtctatgctc attattgtga
      841 tgtgcattgc catctttgcc tgtgtggcct ccacgcttgc ctgggacaga ggctatggaa
      901 cttccctttt aggaggtagt gtaggctacc cttatggagg aagtggcttt ggtagctacg
      961 gaagtggcta tggctatggc tatggttatg gctatggcta cggaggctat acagacccaa
     1021 gagcagcaaa gggcttcatg ttggccatgg ctgccttttg tttcattgcc gcgttggtga
     1081 tctttgttac cagtgttata agatctgaaa tgtccagaac aagaagatac tacttaagtg
     1141 tgataatagt gagtgctatc ctgggcatca tggtgtttat tgccacaatt gtctatataa
     1201 tgggagtgaa cccaactgct cagtcttctg gatctctata tggttcacaa atatatgccc
     1261 tctgcaacca attttataca cctgcagcta ctggactcta cgtggatcag tatttgtatc
     1321 actactgtgt tgtggatccc caggaggcca ttgccattgt actggggttc atgattattg
     1381 tggcttttgc tttaataatt ttctttgctg tgaaaactcg aagaaagatg gacaggtatg
     1441 acaagtccaa tattttgtgg gacaaggaac acatttatga tgagcagccc cccaatgtcg
     1501 aggagtgggt taaaaatgtg tctgcaggca cacaggacgt gccttcaccc ccatctgact
     1561 atgtggaaag agttgacagt cccatggcat actcttccaa tggcaaagtg aatgacaagc
     1621 ggttttatcc agagtcttcc tataaatcca cgccggttcc tgaagtggtt caggagcttc
     1681 cattaacttc gcctgtggat gacttcaggc agcctcgtta cagcagcggt ggtaactttg
     1741 agacaccttc aaaaagagca cctgcaaagg gaagagcagg aaggtcaaag agaacagagc
     1801 aagatcacta tgagacagac tacacaactg gcggcgagtc ctgtgatgag ctggaggagg
     1861 actggatcag ggaatatcca cctatcactt cagatcaaca aagacaactg tacaagagga
     1921 attttgacac tggcctacag gaatacaaga gcttacaatc agaacttgat gagatcaata
     1981 aagaactctc ccgtttggat aaagaattgg atgactatag agaagaaagt gaagagtaca
     2041 tggctgctgc tgatgaatac aatagactga agcaagtgaa gggatctgca gattacaaaa
     2101 gtaagaagaa tcattgcaag cagttaaaga gcaaattgtc acacatcaag aagatggttg
     2161 gagactatga tagacagaaa acaagcggct ccggtagcgg accggtcgcc accatggtga
     2221 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     2281 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2341 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2401 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     2461 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     2521 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     2581 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     2641 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     2701 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     2761 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     2821 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     2881 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
     2941 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3001 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3061 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3121 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     3181 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3241 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3301 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3361 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3421 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3481 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3541 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3601 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3661 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3721 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3781 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3841 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3961 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4021 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4081 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4141 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4201 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4261 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4321 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4381 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4441 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4501 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4561 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4621 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4681 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4741 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4801 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4861 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4921 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4981 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5041 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5101 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5161 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5221 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5281 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5341 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5401 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5461 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5521 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5581 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5641 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5701 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5761 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5821 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5881 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5941 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6001 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6061 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6121 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6181 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6241 ccgt
//
