LOCUS       sequence-403684-        8757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of FLAG-S1PR1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8757)
  TITLE     Constructs expressing FLAG-S1PR1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8757)
  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            "Unpublished"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: pCDH-Flag-hS1pr1_Neo
FEATURES             Location/Qualifiers
     source          1..8757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1807..1923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2013..2216
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2167..2187
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2213..2237
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2294..2317
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        3515..3726
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3739..4007
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4035..4829
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4089..4108)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4699..4718
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    4836..5424
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4889..4909)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5307..5318)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5498..5731
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5803..5924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5840..5859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5894..5913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5964..6099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6132..6148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6132..6148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6145..6167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6156..6172
                     /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(6180..6186,6187..6204,6205..6210))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6225..6246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6363..6380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6534..7122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7293..8084,8085..8153))
                     /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     7916..7935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8154..8258)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8326..8344
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8382..8404)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8504..8523
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8717..8739
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8731..8748
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8732..8748
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
     1921 tttatcgata ctagtattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     1981 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     2041 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     2101 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     2161 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ttatataagc agagctcgtt
     2221 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagat
     2281 tctagacacc atggattaca aggatgacga tgacaaggat atcggtaccg ggcccaccag
     2341 cgtcccgctg gtcaaggccc accgcagctc ggtctctgac tacgtcaact atgatatcat
     2401 cgtccggcat tacaactaca cgggaaagct gaatatcagc gcggacaagg agaacagcat
     2461 taaactgacc tcggtggtgt tcattctcat ctgctgcttt atcatcctgg agaacatctt
     2521 tgtcttgctg accatttgga aaaccaagaa attccaccga cccatgtact attttattgg
     2581 caatctggcc ctctcagacc tgttggcagg agtagcctac acagctaacc tgctcttgtc
     2641 tggggccacc acctacaagc tcactcccgc ccagtggttt ctgcgggaag ggagtatgtt
     2701 tgtggccctg tcagcctccg tgttcagtct cctcgccatc gccattgagc gctatatcac
     2761 aatgctgaaa atgaaactcc acaacgggag caataacttc cgcctcttcc tgctaatcag
     2821 cgcctgctgg gtcatctccc tcatcctggg tggcctgcct atcatgggct ggaactgcat
     2881 cagtgcgctg tccagctgct ccaccgtgct gccgctctac cacaagcact atatcctctt
     2941 ctgcaccacg gtcttcactc tgcttctgct ctccatcgtc attctgtact gcagaatcta
     3001 ctccttggtc aggactcgga gccgccgcct gacgttccgc aagaacattt ccaaggccag
     3061 ccgcagctct gagaagtcgc tggcgctgct caagaccgta attatcgtcc tgagcgtctt
     3121 catcgcctgc tgggcaccgc tcttcatcct gctcctgctg gatgtgggct gcaaggtgaa
     3181 gacctgtgac atcctcttca gagcggagta cttcctggtg ttagctgtgc tcaactccgg
     3241 caccaacccc atcatttaca ctctgaccaa caaggagatg cgtcgggcct tcatccggat
     3301 catgtcctgc tgcaagtgcc cgagcggaga ctctgctggc aaattcaagc gacccatcat
     3361 cgccggcatg gaattcagcc gcagcaaatc ggacaattcc tcccaccccc agaaagacga
     3421 aggggacaac ccagagacca ttatgtcttc tggaaacgtc aactcttctt cctagcggcc
     3481 gcaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     3541 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     3601 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     3661 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     3721 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     3781 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     3841 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     3901 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     3961 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4021 gcgcctacgt cgagatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4081 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4141 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4201 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccgcg acgggcgttc
     4261 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4321 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4381 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4441 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4501 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4561 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4621 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4681 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4741 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4801 tctatcgcct tcttgacgag ttcttctgac tcgacaatca acctctggat tacaaaattt
     4861 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4921 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4981 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5041 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     5101 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     5161 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     5221 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     5281 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     5341 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     5401 tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct
     5461 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     5521 cgaagataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     5581 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5641 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5701 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     5761 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     5821 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5881 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     5941 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6001 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     6061 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagaccaaa
     6121 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6181 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6241 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6301 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6361 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6421 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6481 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6541 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6601 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6661 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6721 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6781 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6841 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6901 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6961 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7021 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7081 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7141 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7201 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7261 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7321 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7381 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7441 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7501 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7561 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7621 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7681 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7741 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7801 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7861 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7921 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7981 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8041 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8101 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8161 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8221 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8281 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8341 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     8401 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     8461 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     8521 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     8581 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     8641 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     8701 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
