LOCUS       sequence-426575-        7361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces human CDC50A-c3xFLAG, a chaperone to phosphatidylserine 
            flippase complexes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7361)
  AUTHORS   Reyes Ballista JM, Hoover AJ, Noble JT, Acciani MD, Miazgowicz KL, 
            Harrison SA, Tabscott GAL, Duncan A, Barnes DN, Jimenez AR, Brindley
            MA
  TITLE     Chikungunya virus release is reduced by TIM-1 receptors through 
            binding of envelope phosphatidylserine.
  JOURNAL   J Virol. 2024 Jul 15:e0077524. doi: 10.1128/jvi.00775-24.
  PUBMED    39007616
REFERENCE   2  (bases 1 to 7361)
  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: "J Virol. 
            2024 Jul 15:e0077524. doi: 10.1128/jvi.00775-24."
COMMENT     Sequence Label: pcDNAintron-CDC50A-c3xFLAG
FEATURES             Location/Qualifiers
     source          1..7361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     815..839
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          939..1511
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1566..1585
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1566..1584
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2843..2908
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2952..2969)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2958..3182
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3228..3656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3315..3334)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3525..3546
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3665..3685)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3670..3999
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3850..3985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3912..3931
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4066..4860
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4120..4139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4730..4749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5034..5155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5071..5090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5125..5144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5204..5220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5204..5220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5217..5239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5228..5244
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5252..5258,5259..5276,5277..5282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5297..5318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5435..5452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5606..6194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5686..5705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6365..7156,7157..7225))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6988..7007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7226..7330)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
      841 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      901 tccgggggat cgatcccccc gatcctgaga acttcagggt gagtttgggg acccttgatt
      961 gttctttctt tttcgctatt gtaaaattca tgttatatgg agggggcaaa gttttcaggg
     1021 tgttgtttag aatgggaaga tgtcccttgt atcaccatgg accctcatga taattttgtt
     1081 tctttcactt tctactctgt tgacaaccat tgtctcctct tattttcttt tcattttctg
     1141 taactttttc gttaaacttt agcttgcatt tgtaacgaat ttttaaattc acttttgttt
     1201 atttgtcaga ttgtaagtac tttctctaat cacttttttt tcaaggcaat cagggtatat
     1261 tatattgtac ttcagcacag ttttagagaa caattgttat aattaaatga taaggtagaa
     1321 tatttctgca tataaattct ggctggcgtg gaaatattct tattggtaga aacaactaca
     1381 tcctggtcat catcctgcct ttctctttat ggttacaatg atatacactg tttgagatga
     1441 ggataaaata ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat gccttcttct
     1501 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     1561 aattgtaata cgactcacta tagggcgaat tcggatccac tagtccagtg tggtggaatt
     1621 ctgcagatct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1681 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg agctcatcga
     1741 tgcatggtac cgatcgatgg cgatgaacta taacgcgaag gatgaagtgg acggtgggcc
     1801 cccgtgtgct ccggggggca ccgcgaagac tcggagaccg gataacacgg ccttcaaaca
     1861 gcaacggctg ccagcttggc agcccatcct tacggctggc acggtgctac ctattttctt
     1921 catcatcggt ctcatcttca ttcccatcgg cattggcatt tttgtcacct ccaacaacat
     1981 ccgcgagatc gagattgatt ataccggaac agagccttcc agtccctgta ataaatgttt
     2041 atctccggat gtgacacctt gcttttgtac cattaacttc acactggaaa agtcatttga
     2101 gggcaacgtg tttatgtatt atggactgtc taatttctat caaaaccatc gtcgttacgt
     2161 gaaatcccga gatgatagtc aactaaatgg agattctagt gctttgctta atcccagtaa
     2221 ggaatgtgaa ccttatcgaa gaaatgaaga caaaccaatt gctccttgtg gagctattgc
     2281 caacagcatg tttaatgata cattagaatt gtttctcatt ggcaatgatt cttatcctat
     2341 acctatcgct ttgaaaaaga aaggtattgc ttggtggaca gataaaaatg tgaaattcag
     2401 aaatccccct ggaggagaca acctggaaga acgatttaaa ggtacaacaa agcctgtgaa
     2461 ctggcttaaa ccagtttaca tgctggattc tgacccagat aataatggat tcataaatga
     2521 ggattttatt gtttggatgc gtactgcagc attacctact tttcgcaagt tgtatcgtct
     2581 tatagaaagg aaaagtgatt tacatccaac attaccagct ggccgatact ctttgaatgt
     2641 cacatacaat taccctgtac attattttga tggacgaaaa cggatgatct tgagcactat
     2701 ttcatggatg ggaggaaaaa atccattttt ggggattgct tacatcgctg ttggatccat
     2761 ctccttcctt ctgggagttg tactgctagt aattaatcat aaatatagaa acagtagtaa
     2821 tacagctgac attaccatta tggactacaa agaccatgac ggtgattata aagatcatga
     2881 tatcgactac aaggatgacg atgacaagta atttctcgag tctagagggc ccgtttaaac
     2941 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3001 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3061 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3121 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3181 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3241 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3301 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3361 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3421 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3481 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3541 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3601 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3661 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3721 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3841 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3901 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3961 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4021 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4081 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4141 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4201 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4261 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4321 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4381 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4441 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4501 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4561 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4621 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4681 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4741 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4801 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4861 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4921 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4981 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5041 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5161 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5221 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5281 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6121 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6181 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6241 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6301 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6361 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6421 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6481 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6541 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6601 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6661 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6721 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6781 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6841 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6901 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6961 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7021 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7081 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7141 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7201 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7261 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7321 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
