LOCUS       sequence-443041-        6244 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 6244)
  TITLE     CRISPR/Cas9 screening reveals a distinct class of pMHC-I binders 
            with precise HLA-peptide recognition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6244)
  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            "Unpublished"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-Delta1
FEATURES             Location/Qualifiers
     source          1..6244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        96..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(574..1162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(654..673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1491..1538
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1563..1582
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1770..2564)
                     /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             complement(1770..2564)
                     /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(1881..1900)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2491..2510
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2599..2956)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2613..2748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2765..2956
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2913..2933
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2958..3062)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3089..3544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3176..3195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3386..3407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3551..3672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3562..3581)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3616..3635
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3794..4507,4508..4510,4511..4513))
                     /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(3839..3860)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4188..4207
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4447..4468
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4510..4519
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(4523..4531,4532..4534,4535..5109)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4696..4715)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     4925..4942
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     promoter        complement(join(6136..6244,1..95))
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6165..6185)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
       61 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
      121 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
      181 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
      241 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
      301 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
      361 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
      421 tgaactaatg accccgtaat tgattactat taataactaa tgcatggcgg taatacggtt
      481 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      541 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      601 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      661 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      721 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      781 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      841 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      901 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      961 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1021 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1081 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1141 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1201 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1261 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaacc
     1321 tgaggctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc cttcacccga
     1381 acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc aatggggtct
     1441 cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga
     1501 cccaacaccg tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg
     1561 tattgtctcc ttccgtgttt cagttagcct ccccctaggg tgggcgaaga actccagcat
     1621 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     1681 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
     1741 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     1801 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     1861 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     1921 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     1981 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc
     2041 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     2101 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     2161 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     2221 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     2281 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     2341 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca
     2401 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     2461 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     2521 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     2581 cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     2641 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     2701 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     2761 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     2821 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     2881 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     2941 gttctttccg cctcaggact cttccttttt caatattatt gaagcattta tcagggttat
     3001 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3061 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     3121 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3181 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3241 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3301 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3361 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3421 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3481 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3541 atttacgcct taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3601 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3661 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3721 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcta
     3781 gagtcgcggc cgcttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     3841 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     3901 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     3961 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     4021 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     4081 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     4141 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     4201 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     4261 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     4321 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     4381 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     4441 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     4501 ccttgctcac catggtggcc atggttgtgg ccatattatc atcgtgtttt tcaaaggaaa
     4561 accacgtccc cgtggttcgg ggggcctaga cgttttttta acctcgacta aacacatgta
     4621 aagcatgtgt accgaggccc cagatcagat cccatacaat ggggtacctt ctgggcatcc
     4681 ttcagcccct tgttgaatac gcttgaggag agccatttga ctctttccac aactatccaa
     4741 ctcacaacgt ggcactgggg ttgtgccgcc tttgcaggtg tatcttatac acgtggcttt
     4801 tggccgcaga ggcacctgtc gccaggtggg gggttccgct gcctgcaaag ggtcgctaca
     4861 gacgttgttt gtcttcaaga agcttccaga ggaactgctt ccttcacgac attcaacaga
     4921 ccttgcattc ctttggcgag aggggaaaga cccctaggaa tgctcgtcaa gaagacaggg
     4981 ccaggtttcc gggccctcac attgccaaaa gacggcaata tggtggaaaa taacatatag
     5041 acaaacgcac accggcctta ttccaagcgg cttcggccag taacgttagg ggggggggag
     5101 ggagaggggc ggatccctaa gtcagcaagc ccatggttac tagcgtccca agcaaacctg
     5161 tcaacgtgaa acacgtgtgc ccagatagaa gacgggtagt acctgaagtg gttccacttc
     5221 ctttatttgg ggttgtttca tgaaaatgct tggttgtcct ggaaacaggt gtactccgtg
     5281 ttgcttgagc atttggtgtg gtggtttttg tggtggtttt ctgagaagtt ggtgagactt
     5341 ctgtagttgg aacatttact gtggtaggtt tctgaactgt tggtgggacc ttggaagtta
     5401 gagattttcc tctgcattca ggtggtgggc cactccactc tccttcatca ttattcacag
     5461 tacaataaat agagtgctct ccaatcatgg tgaatccttt attacatgca tacgttacag
     5521 actgtctata tccataatgg tcacgttccc cttgaattat tccattgtca atttgtggtg
     5581 gtgctggaca ataaatttct ctgcactctg gcaacgggtc actccactgg acagagctgc
     5641 ctgaaataag acaaaaacta gaagtcgagc caaataattt gtaccctgtg ttacacgaga
     5701 aggagatggt tgcaccaaat aatatgccac ctggtacatc aatctgacca tttcgtattt
     5761 ctcccggatt agggcatgat ttctttttac aaaattcgac tgctgtggac cattttaaat
     5821 tctgaaggca agtcagtttt ggtgatagag aaggttctct tctgtaacct ggacggcact
     5881 catattccac aacagtaccg actggaaaat aattctgagt gatataaggc tgtttgaggg
     5941 atgcagaatt tagccttgtt ggcacctcgc agctacgacc ccacacggcc ggcaggcaca
     6001 acagcaccag cagcagcagc cggggcagct ccccgaggag gggcagcgcc gcgggcacgc
     6061 tcggccgcgc gacggtcatg gtggcctcga gatctgagtc cggtagcgct agcggatctg
     6121 acggttcact aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     6181 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     6241 aaac
//
