LOCUS       sequence-443038-        6049 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 6049)
  TITLE     CRISPR/Cas9 screening reveals a distinct class of pMHC-I binders 
            with precise HLA-peptide recognition
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6049)
  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            "Unpublished"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-Delta2
FEATURES             Location/Qualifiers
     source          1..6049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(366..385)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    410..457
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      786..1374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1275..1294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1549..1852
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1853..2056
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2007..2027
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(2888..3462,3463..3465,3466..3474)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3055..3072)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3282..3301
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3478..3487
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3484..3486,3487..3489,3490..4203)
                     /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(3529..3550)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3790..3809)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4137..4158
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4325..4446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4362..4381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4416..4435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4453..4908)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4590..4611)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4802..4821
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4935..5039
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5041..5398
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5041..5232
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5064..5084)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5249..5384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5433..6049,1..178)
                     /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             join(5433..6049,1..178)
                     /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(5487..5506)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
       61 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
      121 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
      181 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
      241 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
      301 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
      361 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
      421 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
      481 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
      541 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
      601 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
      661 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
      721 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
      781 tcttcttgag atcctttttt tctgcgcgta atctgctgca tgcaaacaaa aaaaccaccg
      841 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      901 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      961 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1021 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1081 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1141 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1201 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1261 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1321 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1381 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1441 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     1501 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     1561 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     1621 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1681 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     1741 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1801 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1861 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1921 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     1981 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2041 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat
     2101 ctcgaggcca ccatgaccgt cgcgcggccg agcgtgcccg cggcgctgcc cctcctcggg
     2161 gagctgcccc ggctgctgct gctggtgctg ttgtgcctgc cggccgtgtg gggtaaatca
     2221 tgccctaatc cgggagaaat acgaaatggt cagattgatg taccaggtgg catattattt
     2281 ggtgcaacca tctccttctc atgtaacaca gggtacaaat tatttggctc gacttctagt
     2341 ttttgtctta tttcaggcag ctctgtccag tggagtgacc cgttgccaga gtgcagagaa
     2401 atttattgtc cagcaccacc acaaattgac aatggaataa ttcaagggga acgtgaccat
     2461 tatggatata gacagtctgt aacgtatgca tgtaataaag gattcaccat gattggagaa
     2521 cactctattt attgtactgt gaataatgat gaaggagagt ggagtggccc accacctgaa
     2581 tgcagaggaa aatctctaac ttccaaggtc ccaccaacag ttcagaaacc taccacagta
     2641 aatgttccaa ctacagaagt ctcaccaact tctcagaaaa ccaccacaaa aaccaccaca
     2701 ccaaatgctc aagcaacacg gagtacacct gtttccagga caaccaagca ttttcatgaa
     2761 acaaccccaa ataaaggaag tggaaccact tcaggtacta cccgtcttct atctgggcac
     2821 acgtgtttca cgttgacagg tttgcttggg acgctagtaa ccatgggctt gctgacttag
     2881 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     2941 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     3001 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     3061 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     3121 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     3181 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     3241 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     3301 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     3361 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     3421 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggcc
     3481 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3541 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3601 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3661 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3721 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3781 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3841 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3901 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3961 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4021 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4081 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     4141 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4201 taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4261 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4321 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4441 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4501 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4561 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4621 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4681 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4741 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4801 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4861 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4921 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4981 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5041 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5101 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5161 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5221 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5281 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5341 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5401 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5461 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5521 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5581 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5641 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5701 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5761 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5821 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5881 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5941 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6001 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggatt
//
