LOCUS       CD55_IRES_mGFP_D        6049 bp DNA     circular SYN 27-OCT-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6049)
  AUTHORS   LUMC-Chemical Immunology
  TITLE     Direct Submission
  JOURNAL   Exported May 6, 2024 from SnapGene Viewer 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    609..613
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     primer_bind     612..644
                     /label=Primer 1
                     /note="color: black; sequence: 
                     GactctcgaggccaccATGACCGTCGCGCGGCCGAG"
     primer_bind     complement(1369..1398)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     gactggatccCTAAGTCAGCAAGCCCATGGTTAC"
     misc_feature    1394..1398
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     misc_feature    1400..1986
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="This feature has 3 segments:
                        1: 1400 .. 1974 / #a6acb3
                        2: 1975 .. 1977 / #a6acb3 / ATG
                        3: 1978 .. 1986 / #a6acb3"
     regulatory      1990..1999
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             1996..2715
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1996 .. 1998 / #05fd14
                        2: 1999 .. 2001 / #05fd14 / 1a
                        3: 2002 .. 2715 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2837..2958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(2965..3420)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3447..3551
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        3553..3910
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3761..3896
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3945..4739
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4971..5018
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      5347..5935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 gctagcgcta
      601 ccggactcag atctcgaggc caccatgacc gtcgcgcggc cgagcgtgcc cgcggcgctg
      661 cccctcctcg gggagctgcc ccggctgctg ctgctggtgc tgttgtgcct gccggccgtg
      721 tggggtaaat catgccctaa tccgggagaa atacgaaatg gtcagattga tgtaccaggt
      781 ggcatattat ttggtgcaac catctccttc tcatgtaaca cagggtacaa attatttggc
      841 tcgacttcta gtttttgtct tatttcaggc agctctgtcc agtggagtga cccgttgcca
      901 gagtgcagag aaatttattg tccagcacca ccacaaattg acaatggaat aattcaaggg
      961 gaacgtgacc attatggata tagacagtct gtaacgtatg catgtaataa aggattcacc
     1021 atgattggag agcactctat ttattgtact gtgaataatg atgaaggaga gtggagtggc
     1081 ccaccacctg aatgcagagg aaaatctcta acttccaagg tcccaccaac agttcagaaa
     1141 cctaccacag taaatgttcc aactacagaa gtctcaccaa cttctcagaa aaccaccaca
     1201 aaaaccacca caccaaatgc tcaagcaaca cggagtacac ctgtttccag gacaaccaag
     1261 cattttcatg aaacaacccc aaataaagga agtggaacca cttcaggtac tacccgtctt
     1321 ctatctgggc acacgtgttt cacgttgaca ggtttgcttg ggacgctagt aaccatgggc
     1381 ttgctgactt agggatccgc ccctctccct cccccccccc taacgttact ggccgaagcc
     1441 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     1501 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     1561 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     1621 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     1681 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     1741 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     1801 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     1861 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     1921 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     1981 acaaccatgg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2041 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2101 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2161 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2221 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2281 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2341 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2401 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2461 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2521 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2581 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     2641 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2701 gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2761 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2821 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2881 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2941 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3001 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3061 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3121 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3181 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3241 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3301 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3361 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3421 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3481 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3541 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3601 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3661 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3721 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3781 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3841 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3901 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3961 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4021 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4081 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4141 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4201 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4261 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4321 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4381 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4441 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4501 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4561 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4621 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4681 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4741 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4801 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4861 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4921 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4981 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5041 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5101 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5161 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5461 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6001 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
