LOCUS       sequence-235388-        5572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin Capping Protein , C1 Cloning Vector, 
            Excitation:487, Emission:509.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5572)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 5572)
  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: mEmerlad-MCPB2-C-18
FEATURES             Location/Qualifiers
     source          1..5572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      631..640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(637..639,640..642,643..1353)
                     /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(682..703)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(943..962)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1290..1311
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2384..2505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2421..2440)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2475..2494
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2512..2967)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2649..2670)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2861..2880
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2994..3098
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3100..3457
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3100..3291
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3123..3143)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3308..3443
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3492..4286
                     /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             3492..4286
                     /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(3546..3565)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4156..4175
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4474..4493)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4518..4565
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4894..5482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5383..5402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      601 gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
      661 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      721 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      781 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt gacctacggc
      841 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      901 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      961 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1021 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1081 cacaaggtct atatcaccgc cgacaagcag aagaacggca tcaaggtgaa cttcaagacc
     1141 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1201 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     1261 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1321 gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctcg agagtccagt
     1381 ggatattcgt ctcggtcggg tggaagcagc gatcagcagc tggactgcgc cttggacctg
     1441 atgaggcgcc tgcctccaca gcagattgag aagaacctca gcgatctgat cgacctggtc
     1501 cccagtctgt gtgaagatct cctgtcatct gttgaccagc ccctgaaaat tgccagagac
     1561 aaggtggtgg gcaaggatta ccttttgtgt gactacaaca gagacgggga ctcctatagg
     1621 tcaccgtgga gtaacaagta tgaccctcct ttggaagatg gggccatgcc atctgctcgg
     1681 ctcagaaagc tggaggtaga ggccaacaat gccttcgacc aataccgaga cctgtatttt
     1741 gaaggtgggg tctcatcagt ctacctctgg gatcttgatc atggctttgc tggagtgatc
     1801 ctcataaaga aagctggaga tggatccaag aagatcaaag gctgctggga ttccatccac
     1861 gtggtggaag tgcaggagaa gtccagcggc cgtactgccc attacaagtt gacctccacg
     1921 gtgatgctat ggctgcaaac caacaaatcc ggctcgggca ccatgaacct gggaggcagc
     1981 ctaaccagac agatggagaa agacgaaact gtgagtgact gttccccaca catagccaac
     2041 atcgggcgcc tggtggagga catggaaaac aaaatccgaa gcacgctgaa tgagatctac
     2101 tttggaaaaa caaaggacat cgtcaacggg ctgaggtctg tgcagacgtt tgcagacaaa
     2161 tcaaagcaag aagcgcttaa gaacgacctg gtggaggcct tgaagagaaa gcagcagtgt
     2221 tgagaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat
     2281 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2341 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2401 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2461 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     2521 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2581 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2641 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2701 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2761 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2821 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2881 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2941 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3001 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3061 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3121 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3181 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3301 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3361 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3421 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3481 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3541 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3601 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3661 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3721 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3781 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3841 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3901 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3961 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4021 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4081 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4141 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4201 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4261 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4321 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4381 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4441 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4501 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4561 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4621 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4681 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4741 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4801 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4861 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5041 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5521 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tg
//
