LOCUS       sequence-446054-        8267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8267)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 8267)
  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-Integrin-Alpha2-N-18
FEATURES             Location/Qualifiers
     source          1..8267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4156..4165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4162..4872
                     /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(4307..4325)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4575..4594
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4995..5116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5032..5051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5086..5105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5123..5578)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5260..5281)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5472..5491
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5605..5709
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5711..6068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5711..5902
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5734..5754)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5919..6054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6103..6897
                     /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             6103..6897
                     /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(6157..6176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6767..6786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7085..7104)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7129..7176
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7505..8093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7994..8013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagac cgccatgggg ccagaacgga caggggccgc gccgctgccg
      601 ctgctgctgg tgttagcgct cagtcaaggc attttaaatt gttgtttggc ctacaatgtt
      661 ggtctcccag aagcaaaaat attttccggt ccttcaagtg aacagtttgg ctatgcagtg
      721 cagcagttta taaatccaaa aggcaactgg ttactggttg gttcaccctg gagtggcttt
      781 cctgagaacc gaatgggaga tgtgtataaa tgtcctgttg acctatccac tgccacatgt
      841 gaaaaactaa atttgcaaac ttcaacaagc attccaaatg ttactgagat gaaaaccaac
      901 atgagcctcg gcttgatcct caccaggaac atgggaactg gaggttttct cacatgtggt
      961 cctctgtggg cacagcaatg tgggaatcag tattacacaa cgggtgtgtg ttctgacatc
     1021 agtcctgatt ttcagctctc agccagcttc tcacctgcaa ctcagccctg cccttccctc
     1081 atagatgttg tggttgtgtg tgatgaatca aatagtattt atccttggga tgcagtaaag
     1141 aattttttgg aaaaatttgt acaaggcctg gatataggcc ccacaaagac acaggtgggg
     1201 ttaattcagt atgccaataa tccaagagtt gtgtttaact tgaacacata taaaaccaaa
     1261 gaagaaatga ttgtagcaac atcccagaca tcccaatatg gtggggacct cacaaacaca
     1321 tttggagcaa ttcaatatgc aagaaaatat gcttattcag cagcttctgg tgggcgacga
     1381 agtgctacaa aagtaatggt agttgtaact gacggtgaat cacatgatgg ttcaatgttg
     1441 aaagctgtga ttgatcaatg caaccatgac aatatactga ggtttggcat agcagttctt
     1501 gggtacttaa acagaaacgc ccttgatact aaaaatttaa taaaagaaat aaaagcaatc
     1561 gctagtattc caacagaaag atactttttc aatgtgtctg atgaagcagc tctactagaa
     1621 aaggctggga cattaggaga acaaattttc agcattgaag gtactgttca aggaggagac
     1681 aactttcaga tggaaatgtc acaagtggga ttcagtgcag attactcttc tcaaaatgat
     1741 attctgatgc tgggtgcagt gggagctttt ggctggagtg ggaccattgt ccagaagaca
     1801 tctcatggcc atttgatctt tcctaaacaa gcctttgacc aaattctgca ggacagaaat
     1861 cacagttcat atttaggtta ctctgtggct gcaatttcta ctggagaaag cactcacttt
     1921 gttgctggtg ctcctcgggc aaattatacc ggccagatag tgctatatag tgtgaatgag
     1981 aatggcaata tcacggttat tcaggctcac cgaggtgacc agattggctc ctattttggt
     2041 agtgtgctgt gttcagttga tgtggataaa gacaccatta cagacgtgct cttggtaggt
     2101 gcaccaatgt acatgagtga cctaaagaaa gaggaaggaa gagtctacct gtttactatc
     2161 aaagagggca ttttgggtca gcaccaattt cttgaaggcc ccgagggcat tgaaaacact
     2221 cgatttggtt cagcaattgc agctctttca gacatcaaca tggatggctt taatgatgtg
     2281 attgttggtt caccactaga aaatcagaat tctggagctg tatacattta caatggtcat
     2341 cagggcacta tccgcacaaa gtattcccag aaaatcttgg gatccgatgg agcctttagg
     2401 agccatctcc agtactttgg gaggtccttg gatggctatg gagatttaaa tggggattcc
     2461 atcaccgatg tgtctattgg tgcctttgga caagtggttc aactctggtc acaaagtatt
     2521 gctgatgtag ctatagaagc ttcattcaca ccagaaaaaa tcactttggt caacaagaat
     2581 gctcagataa ttctcaaact ctgcttcagt gcaaagttca gacctactaa gcaaaacaat
     2641 caagtggcca ttgtatataa catcacactt gatgcagatg gattttcatc cagagtaacc
     2701 tccagggggt tatttaaaga aaacaatgaa aggtgcctgc agaagaatat ggtagtaaat
     2761 caagcacaga gttgccccga gcacatcatt tatatacagg agccctctga tgttgtcaac
     2821 tctttggatt tgcgtgtgga catcagtctg gaaaaccctg gcactagccc tgcccttgaa
     2881 gcctattctg agactgccaa ggtcttcagt attcctttcc acaaagactg tggtgaggac
     2941 ggactttgca tttctgatct agtcctagat gtccgacaaa taccagctgc tcaagaacaa
     3001 ccctttattg tcagcaacca aaacaaaagg ttaacatttt cagtaacgct gaaaaataaa
     3061 agggaaagtg catacaacac tggaattgtt gttgattttt cagaaaactt gttttttgca
     3121 tcattctccc tgccggttga tgggacagaa gtaacatgcc aggtggctgc atctcagaag
     3181 tctgttgcct gcgatgtagg ctaccctgct ttaaagagag aacaacaggt gacttttact
     3241 attaactttg acttcaatct tcaaaacctt cagaatcagg cgtctctcag tttccaagcc
     3301 ttaagtgaaa gccaagaaga aaacaaggct gataatttgg tcaacctcaa aattcctctc
     3361 ctgtatgatg ctgaaattca cttaacaaga tctaccaaca taaattttta tgaaatctct
     3421 tcggatggga atgttccttc aatcgtgcac agttttgaag atgttggtcc aaaattcatc
     3481 ttctccctga aggtaacaac aggaagtgtt ccagtaagca tggcaactgt aatcatccac
     3541 atccctcagt ataccaaaga aaagaaccca ctgatgtacc taactggggt gcaaacagac
     3601 aaggctggtg acatcagttg taatgcagat atcaatccac tgaaaatagg acaaacatct
     3661 tcttctgtat ctttcaaaag tgaaaatttc aggcacacca aagaattgaa ctgcagaact
     3721 gcttcctgta gtaatgttac ctgctggttg aaagacgttc acatgaaagg agaatacttt
     3781 gttaatgtga ctaccagaat ttggaacggg acttttgcat catcaacgtt ccagacagta
     3841 cagctaacgg cagctgcaga aatcaacacc tataaccctg agatatatgt gattgaagat
     3901 aacactgtta cgattcccct gatgataatg aaacctgatg agaaagccga agtaccaaca
     3961 ggagttataa taggaagtat aattgctgga atccttttgc tgttagctct ggttgcaatt
     4021 ttatggaagc tcggcttctt caaaagaaaa tatgaaaaga tgaccaaaaa tccagatgag
     4081 attgatgaga ccacagagct cagtagcggc agcgcaggcg gaagcggggt accgcgggcc
     4141 cgggatccac cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc
     4201 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     4261 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     4321 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     4381 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     4441 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     4501 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     4561 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     4621 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     4681 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     4741 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     4801 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     4861 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     4921 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     4981 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5041 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5101 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     5161 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5221 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5281 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5341 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5401 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5461 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5521 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     5581 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5641 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5701 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     5761 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5821 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5881 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5941 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6001 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6061 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     6121 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     6181 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     6241 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     6301 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     6361 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     6421 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     6481 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     6541 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     6601 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     6661 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6721 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6781 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6841 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6901 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6961 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     7021 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     7081 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     7141 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     7201 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     7261 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     7321 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     7381 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7441 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7501 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7561 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7621 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7681 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7741 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7801 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7861 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7921 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7981 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8041 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8101 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8161 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     8221 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
