LOCUS       sequence-443039-        6427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CD55 variant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6427)
  TITLE     CRISPR/Cas9 screening reveals a distinct class of pMHC-I binders 
            with precise HLA-peptide recognition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6427)
  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            "Unpublished"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: CD55-full length
FEATURES             Location/Qualifiers
     source          1..6427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..70
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1309..1883,1884..1886,1887..1895)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1476..1493)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1703..1722
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1899..1908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1905..1907,1908..1910,1911..2624)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1950..1971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2211..2230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2558..2579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2746..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2837..2856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2874..3329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3011..3032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3223..3242
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3356..3460
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3462..3819
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3462..3653
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3485..3505)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3670..3805
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3854..4648
                     /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             3854..4648
                     /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(3908..3927)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4518..4537
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4836..4855)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4880..4927
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5256..5844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5745..5764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6019..6322
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6323..6427,1..99)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
       61 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      121 cgctagcgct accggactca gatctcgagg ccaccatgac cgtcgcgcgg ccgagcgtgc
      181 ccgcggcgct gcccctcctc ggggagctgc cccggctgct gctgctggtg ctgttgtgcc
      241 tgccggccgt gtggggtgac tgtggccttc ccccagatgt acctaatgcc cagccagctt
      301 tggaaggccg tacaagtttt cccgaggata ctgtaataac gtacaaatgt gaagaaagct
      361 ttgtgaaaat tcctggcgag aaggactcag tgatctgcct taagggcagt caatggtcag
      421 atattgaaga gttctgcaat cgtagctgcg aggtgccaac aaggctaaat tctgcatccc
      481 tcaaacagcc ttatatcact cagaattatt ttccagtcgg tactgttgtg gaatatgagt
      541 gccgtccagg ttacagaaga gaaccttctc tatcaccaaa actaacttgc cttcagaatt
      601 taaaatggtc cacagcagtc gaattttgta aaaagaaatc atgccctaat ccgggagaaa
      661 tacgaaatgg tcagattgat gtaccaggtg gcatattatt tggtgcaacc atctccttct
      721 catgtaacac agggtacaaa ttatttggct cgacttctag tttttgtctt atttcaggca
      781 gctctgtcca gtggagtgac ccgttgccag agtgcagaga aatttattgt ccagcaccac
      841 cacaaattga caatggaata attcaagggg aacgtgacca ttatggatat agacagtctg
      901 taacgtatgc atgtaataaa ggattcacca tgattggaga gcactctatt tattgtactg
      961 tgaataatga tgaaggagag tggagtggcc caccacctga atgcagagga aaatctctaa
     1021 cttccaaggt cccaccaaca gttcagaaac ctaccacagt aaatgttcca actacagaag
     1081 tctcaccaac ttctcagaaa accaccacaa aaaccaccac accaaatgct caagcaacac
     1141 ggagtacacc tgtttccagg acaaccaagc attttcatga aacaacccca aataaaggaa
     1201 gtggaaccac ttcaggtact acccgtcttc tatctgggca cacgtgtttc acgttgacag
     1261 gtttgcttgg gacgctagta accatgggct tgctgactta gggatccgcc cctctccctc
     1321 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1381 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     1441 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     1501 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     1561 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     1621 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     1681 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     1741 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca catgctttac
     1801 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     1861 cctttgaaaa acacgatgat aatatggcca caaccatggc caccatggtg agcaagggcg
     1921 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1981 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2041 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2101 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2161 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2221 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2281 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2341 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2401 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2461 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2521 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2581 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcggcc gcgactctag
     2641 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2701 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2761 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2821 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2881 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2941 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3001 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3061 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3121 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3181 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3241 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3301 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3361 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3421 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3481 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3541 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3661 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3721 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3781 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3841 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3901 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3961 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4021 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4081 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4141 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4201 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4261 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4321 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4381 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4441 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4501 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4561 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4621 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4681 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4741 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4801 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4861 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4921 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4981 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5041 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5101 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5161 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5221 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5281 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5341 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5401 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5461 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5521 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5581 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5641 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5701 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5761 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5821 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5881 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5941 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     6001 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     6061 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     6121 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     6181 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     6241 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     6301 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     6361 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     6421 ttggcac
//
