LOCUS       sequence-456544-        6488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Endoplasmic Reticulum, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 6488)
  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-Calnexin-N-14
FEATURES             Location/Qualifiers
     source          1..6488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..31)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    849..896
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     921..940
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1128..1922)
                     /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             complement(1128..1922)
                     /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(1239..1258)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1849..1868
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1957..2314)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1971..2106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2123..2314
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2271..2291
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2316..2420)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2447..2902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2534..2553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2744..2765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2909..3030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2920..2939)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2974..2993
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3153..3863)
                     /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(3431..3450)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3700..3718
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3860..3869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5738..5941)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5767..5787)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5942..6245
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(join(6420..6488,1..520))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
       61 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      541 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      601 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      661 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
      721 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
      781 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
      841 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
      901 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
      961 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     1021 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     1081 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     1141 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     1201 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     1261 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     1321 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     1381 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     1441 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     1501 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     1561 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     1621 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     1681 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     1741 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     1801 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     1861 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     1921 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     1981 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     2041 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     2101 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     2161 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     2221 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     2281 acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca atattattga
     2341 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2401 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     2461 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2521 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2581 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2641 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2701 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2761 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2821 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2881 aaaatattaa cgcttacaat ttacgcctta agatacattg atgagtttgg acaaaccaca
     2941 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3001 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3061 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     3121 atggctgatt atgatctaga gtcgcggccg ctttacttgt acagctcgtc catgccgccg
     3181 gtggagtggc ggccctcggc gcgttcgtac tgttccacga tggtgtagtc ctcgttgtgg
     3241 gaggtgatgt ccaacttgat gttgacgttg taggcgccgg gcagctgcac gggcttcttg
     3301 gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc cgtccttcag cttcagcctc
     3361 tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca tccgctcgga ggaggcctcc
     3421 cagcccatgg tcttcttctg cattacgggg ccgtcggagg ggaagttggt gccgcgcagc
     3481 ttcaccttgt agatgaactc gccgtcctgc agggaggagt cctgggtcac ggtcaccacg
     3541 ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa ggacagcttc
     3601 aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct tggagccgta catgaactga
     3661 ggggacagga tgtcccaggc gaagggcagg gggccaccct tggtcacctt cagcttggcg
     3721 gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg
     3781 ttcacggagc cctccatgtg caccttgaag cgcatgaact ccttgatgat ggccatgtta
     3841 tcctcctcgc ccttgctcac catggtggcg accggtccag atccgctgcc gcctgacgca
     3901 cccccctctc ttcgtggctt tctgtttctt ggtgatctgt tcaaaatttc atcctcctct
     3961 gctttaggtt ttctgtcttc ctcctcttga ctgacagtgc caccatcttc ttcagcatca
     4021 cttttctgtt tctcttcaag tttctcttct ccttcctcct cctcatctcc cttgtccttt
     4081 tcctcttcct tctcttcttc ctcttccttc acatccggtt gaggtgcatc agttttctta
     4141 tactccatac cactggtctg tttctttcca gaacagcaga agaggataac caggaacaca
     4201 ggaagggcta cagttagaat atagactacc cacagccacg ggcgctcttc agctgcctcg
     4261 ttcatctgcc ccacaacgcc tggctcagca gccccatcag cagctttctt caggccccat
     4321 ccatcattgg cccaatcatc aactattctt cgatcagcac aaatgataaa gttgtcaaaa
     4381 aaaatgtcag aggtcatgga ccacagctcc aaaccaatag cactaaaagg agtcattctg
     4441 aaaggttcca gatcttcaaa gaaatctgga tttggtattt tcctgggttt ccagattccc
     4501 tggtaactgg gattgtcaat cataggaggc ttccatttgc ctttataatt ggggttgtca
     4561 atcacaggtc gctgccagac accacatcca ggagctgact cacatctagg gttggcaatc
     4621 tgaggagcct cccattctcc atccatgtct tcatcccaat cctcaggttt ctctgcgtct
     4681 ggatcaggta cgtactcagg ctcatcatct aaccagcctt cgggttttgt ggcctcttca
     4741 tctggaatct tagcaggggc atcttcatcc cagtcatctg gcttgacagc ttctggatct
     4801 gggatttttg gtctttcatc ccaatcctcg ggcttccggt cttctgggtc ctcaatttca
     4861 cgtgaaggat ttacaggagg agtcatgtca ttgagcagat ttccactatt caccacagat
     4921 tggtcaacca gtatttcaaa actattatct ggattcaaga ttagtgtgta aagatgtgtt
     4981 ttcttatcag taaaataggt cttcagatct gcatctggcc tcttagcatg tttttcttca
     5041 tagatacccg ttttggggtt tttgtgtcgg aagatgaagt gcagtttata gtcctctcca
     5101 catttatctg gaccaaacat aatcgtataa ggggtcttgt catggaactg atccaggttg
     5161 agttctggtg ttttagaaag cagtttcaca taggcaccac cacattctat tccattttgg
     5221 aaattaacct catactgaac aatgagaggc ttggtgtcaa acaggaaggg cttgttcagt
     5281 ttagcagaga tggcatgatg cttggcccga gacatcaaca caagtccttt atcacctgga
     5341 agctttgact ccttcatttc ctctacctcc cactttccat catatttggc aatttcatca
     5401 tcggtatcgt ctttcttggc tttggataaa atccaccctg acagagttcc tctgtcaaaa
     5461 gaatcagcaa aatatacttc ccctgttgga actggagctt tgtaagtaac cttgggagat
     5521 gaaggaggag cagtggtatc tggttttgag tcttctacct cttcaatgac atcgtcaagg
     5581 tcatcctcaa tatcaatcac atcatcatca tgtccatcat gagcctcaac aatagcagtt
     5641 ccaagcacca gtaacataca cagcaaccac ttcccttcca tggcggtctc gagatctgag
     5701 tccggtagcg ctagcggatc tgacggttca ctaaaccagc tctgcttata tagacctccc
     5761 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     5821 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     5881 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     5941 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     6001 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     6061 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     6121 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     6181 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     6241 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     6301 aactaatgca tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6361 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6421 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6481 cgaaaccc
//
