LOCUS       sequence-70482-e        5606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AmCyan while the mCherry gene has been mutated.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5606)
  TITLE     AmCyan-P2A-Xerry
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5606)
  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            "Unpublished"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: AmCyan-P2A-Xerry
FEATURES             Location/Qualifiers
     source          1..5606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             630..650
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             654..674
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             678..698
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             714..1400
                     /codon_start=1
                     /product="Anemonia majano cyan fluorescent protein"
                     /label=AmCyan
                     /translation="MALSNKFIGDDMKMTYHMDGCVNGHYFTVKGEGSGKPYEGTQTST
                     FKVTMANGGPHAFSFDILSTVFMYGNRCFTAYPTSMPDYFKQAFPDGMSYERTFTYEDG
                     GVATASWEISLKGNCFEHKSTFHGVNFPADGPVMAKMTTGWDPSFEKMTVCDGILKGDV
                     TAFLMLQGGGNYRCQFHTSYKTKKPVTMPPNHAVEHRIARTDLDKGGNSVQLTEHAVAH
                     ITSVVPF"
     CDS             1422..1478
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(1636..1654)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1903..1922
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2214..2270
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2394..2515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2431..2450)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2485..2504
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2522..2977)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2659..2680)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2871..2890
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3004..3108
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3110..3467
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3110..3301
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3133..3153)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3318..3453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3502..4296
                     /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             3502..4296
                     /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(3556..3575)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4166..4185
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4484..4503)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4528..4575
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4904..5492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5393..5412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc cgtctccgta
      601 cgatggccac aaccatggtg cacgtggatc caaaaaagaa gagaaaggta gatccaaaaa
      661 agaagagaaa ggtagatcca aaaaagaaga gaaaggtaca cgtgtccacg cgtatggctc
      721 tttcaaacaa gtttatcgga gatgacatga aaatgaccta ccatatggat ggctgtgtca
      781 atgggcatta ctttaccgtc aaaggtgaag gcagcgggaa gccatacgaa gggacgcaga
      841 cctcgacttt taaagtcacc atggccaacg gtgggcccca tgcattctcc tttgacatac
      901 tatctacagt gttcatgtat ggaaatcgat gctttactgc gtatcctacc agtatgcccg
      961 actatttcaa acaagcattt cctgacggaa tgtcatatga aaggactttt acctatgaag
     1021 atggaggagt tgctacagcc agttgggaaa taagccttaa aggcaactgc tttgagcaca
     1081 aatccacgtt tcatggagtg aactttcctg ctgatggacc tgtgatggcg aagatgacaa
     1141 ctggttggga cccatctttt gagaaaatga ctgtctgcga tggaatattg aagggtgatg
     1201 tcaccgcgtt cctcatgctg caaggaggtg gcaattacag atgccaattc cacacttctt
     1261 acaagacaaa aaaaccggtg acgatgccac caaaccatgc ggtggaacat cgcattgcga
     1321 ggaccgacct tgacaaaggt ggcaacagtg ttcagctgac ggagcacgct gttgcacata
     1381 taacctctgt tgtccctttc gcgcgcgaga cgggaagcgg agctactaac ttcagcctgc
     1441 tgaagcaggc tggcgacgtg gaggagaacc ctggacctgg tctccggccg atggtgagca
     1501 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     1561 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1621 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     1681 acatcctgtc ccctcagttc atgtaggctc caaggcctac gtgaagcacc ccgccgacat
     1741 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1801 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1861 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1921 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1981 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     2041 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     2101 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     2161 ccgccactcc accggcggca tggacgagct gtacaagtcc ggaaactagt ctcagatctc
     2221 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     2281 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2341 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2401 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2461 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     2521 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2581 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2641 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2701 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2761 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2821 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2881 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2941 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3001 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3061 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3121 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3181 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3301 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3361 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3421 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3481 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3541 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3601 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3661 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3721 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3781 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3841 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3901 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3961 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4021 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4081 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4141 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4201 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4261 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4321 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4381 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4441 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4501 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4561 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4621 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4681 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4741 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4801 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4861 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4921 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4981 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5041 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5101 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5161 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5221 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5281 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5341 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5401 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5461 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5521 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5581 tggataaccg tattaccgcc atgcat
//
