LOCUS       CD55_IRES_mGFP_F        6427 bp DNA     circular SYN 06-MAY-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6427)
  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..6427
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     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"
     CDS             625..1770
                     /codon_start=1
                     /label=CD55 FL
                     /note="color: #993366"
                     /translation="MTVARPSVPAALPLLGELPRLLLLVLLCLPAVWGDCGLPPDVPNA
                     QPALEGRTSFPEDTVITYKCEESFVKIPGEKDSVICLKGSQWSDIEEFCNRSCEVPTRL
                     NSASLKQPYITQNYFPVGTVVEYECRPGYRREPSLSPKLTCLQNLKWSTAVEFCKKKSC
                     PNPGEIRNGQIDVPGGILFGATISFSCNTGYKLFGSTSSFCLISGSSVQWSDPLPECRE
                     IYCPAPPQIDNGIIQGERDHYGYRQSVTYACNKGFTMIGEHSIYCTVNNDEGEWSGPPP
                     ECRGKSLTSKVPPTVQKPTTVNVPTTEVSPTSQKTTTKTTTPNAQATRSTPVSRTTKHF
                     HETTPNKGSGTTSGTTRLLSGHTCFTLTGLLGTLVTMGLLT"
     primer_bind     complement(1747..1776)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     gactggatccCTAAGTCAGCAAGCCCATGGTTAC"
     misc_feature    1772..1776
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     misc_feature    1778..2364
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="This feature has 3 segments:
                        1: 1778 .. 2352 / #a6acb3
                        2: 2353 .. 2355 / #a6acb3 / ATG
                        3: 2356 .. 2364 / #a6acb3"
     regulatory      2368..2377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             2374..3093
                     /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: 2374 .. 2376 / #05fd14
                        2: 2377 .. 2379 / #05fd14 / 1a
                        3: 2380 .. 3093 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3215..3336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(3343..3798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3825..3929
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        3931..4288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4139..4274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4323..5117
                     /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    5349..5396
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      5725..6313
                     /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 tggggtgact gtggccttcc cccagatgta cctaatgccc agccagcttt ggaaggccgt
      781 acaagttttc ccgaggatac tgtaataacg tacaaatgtg aagaaagctt tgtgaaaatt
      841 cctggcgaga aggactcagt gatctgcctt aagggcagtc aatggtcaga tattgaagag
      901 ttctgcaatc gtagctgcga ggtgccaaca aggctaaatt ctgcatccct caaacagcct
      961 tatatcactc agaattattt tccagtcggt actgttgtgg aatatgagtg ccgtccaggt
     1021 tacagaagag aaccttctct atcaccaaaa ctaacttgcc ttcagaattt aaaatggtcc
     1081 acagcagtcg aattttgtaa aaagaaatca tgccctaatc cgggagaaat acgaaatggt
     1141 cagattgatg taccaggtgg catattattt ggtgcaacca tctccttctc atgtaacaca
     1201 gggtacaaat tatttggctc gacttctagt ttttgtctta tttcaggcag ctctgtccag
     1261 tggagtgacc cgttgccaga gtgcagagaa atttattgtc cagcaccacc acaaattgac
     1321 aatggaataa ttcaagggga acgtgaccat tatggatata gacagtctgt aacgtatgca
     1381 tgtaataaag gattcaccat gattggagag cactctattt attgtactgt gaataatgat
     1441 gaaggagagt ggagtggccc accacctgaa tgcagaggaa aatctctaac ttccaaggtc
     1501 ccaccaacag ttcagaaacc taccacagta aatgttccaa ctacagaagt ctcaccaact
     1561 tctcagaaaa ccaccacaaa aaccaccaca ccaaatgctc aagcaacacg gagtacacct
     1621 gtttccagga caaccaagca ttttcatgaa acaaccccaa ataaaggaag tggaaccact
     1681 tcaggtacta cccgtcttct atctgggcac acgtgtttca cgttgacagg tttgcttggg
     1741 acgctagtaa ccatgggctt gctgacttag ggatccgccc ctctccctcc ccccccccta
     1801 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     1861 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     1921 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     1981 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     2041 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     2101 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     2161 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     2221 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     2281 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     2341 cacgatgata atatggccac aaccatggcc accatggtga gcaagggcga ggagctgttc
     2401 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2461 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2521 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2581 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2641 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2701 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2761 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2821 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2881 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2941 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     3001 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3061 atcactctcg gcatggacga gctgtacaag taagcggccg cgactctaga tcataatcag
     3121 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3181 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3241 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3301 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3361 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3421 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3481 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3541 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3601 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3661 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3721 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3781 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3841 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3901 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3961 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4021 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4141 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4201 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4261 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4321 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4381 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4441 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4501 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4561 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4621 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4681 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4741 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4801 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4861 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4921 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4981 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5041 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5101 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5161 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5221 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5281 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5341 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5401 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5461 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5521 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5581 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5641 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6421 catgcat
//
