LOCUS       sequence-436028-        7344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human FLVCR1/FLVCR1a with C-terminal 3x-FLAG cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7344)
  AUTHORS   Kenny TC, Khan A, Son Y, Yue L, Heissel S, Sharma A, Pasolli HA, Liu
            Y, Gamazon ER, Alwaseem H, Hite RK, Birsoy K
  TITLE     Integrative genetic analysis identifies FLVCR1 as a plasma-membrane 
            choline transporter in mammals.
  JOURNAL   Cell Metab. 2023 Jun 6;35(6):1057-1071.e12. doi: 
            10.1016/j.cmet.2023.04.003. Epub 2023 Apr 25.
  PUBMED    37100056
REFERENCE   2  (bases 1 to 7344)
  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"; doi: 
            "10.1016/j.cmet.2023.04.003"; journalName: "Cell Metab"; date: 
            "2023-06-6- 6"; volume: "35"; issue: "6"; pages: "1057-1071.e12"
COMMENT     Sequence Label: pMXS-IRES-Blast FLVCR1/FLVCR1a-3X FLAG
FEATURES             Location/Qualifiers
     source          1..7344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             21..614
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    677..1034
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1093..1509
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1439..1461
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1477..1493
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1519..1893
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1831..1847
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             3602..3667
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3695..4269,4270..4272,4273..4281)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3862..3879)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4089..4108
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4282..4677
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4848..5317
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5447..5464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5618..6206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5698..5717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6370..7161,7162..7230))
                     /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     6993..7012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7231..7335)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgcat aatgaaagac cccacctgta ggtttggcaa gctagcttaa
       61 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagaaaa gttcagatca
      121 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      181 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
      241 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
      301 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
      361 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
      421 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
      481 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
      541 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
      601 gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga
      661 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
      721 ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc
      781 ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg agacgtccca
      841 gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga tcgttttgga
      901 ctctttggtg caccccccta ataggaggga tatgtggttc tggtaggaga cgagaaccta
      961 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg
     1021 cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     1081 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     1141 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     1201 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     1261 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     1321 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     1381 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     1441 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     1501 ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc ccttgtaaac
     1561 ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc tcacttacag
     1621 gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta ccaagaacaa
     1681 ctggaccgac cggtggtacc tcacccttac cgagtcggcg acacagtgtg ggtccgccga
     1741 caccagacta agaacctaga acctcgctgg aaaggacctt acacagtcct gctgaccacc
     1801 cccaccgccc tcaaagtaga cggcatcgca gcttggatac acgccgccca cgtgaaggct
     1861 gccgaccccg ggggtggacc atcctctaga ctgccggatc tagctagtta attaaatggc
     1921 ccgccccgat gacgaagagg gagctgccgt agctcctggc catccactgg ccaaggggta
     1981 tctgcctctc ccacggggtg cccctgtggg aaaagaatcc gtcgaacttc aaaatggccc
     2041 taaggccgga acttttcctg ttaacggcgc gcctcgtgat tctctggcag ctgctagcgg
     2101 tgtattgggt ggaccacaaa ccccacttgc acccgaagaa gaaacacaag ccagattgct
     2161 gccagccggc gcgggcgccg aaacccccgg cgccgaatca tcaccacttc ctctgacagc
     2221 cctgtctcct agacgatttg tagtcctgct catatttagt ctttactctc tcgtgaacgc
     2281 attccaatgg atacagtatt caataatctc aaatgtgttt gaaggatttt atggcgtgac
     2341 tctgttgcat atagattggc ttagcatggt ctatatgctc gcttatgttc ctctgatatt
     2401 tccagctaca tggcttctcg atactagggg tctccgcctt acagcactcc tcgggtcagg
     2461 tctgaattgt ctcggagcat ggataaaatg tggatccgtc caacaacacc tgttttgggt
     2521 gacgatgctc gggcaatgtc tttgttcagt tgcacaagtc tttattctcg ggctgccatc
     2581 aagaattgct tctgtctggt tcggccctaa ggaagtaagc acggcctgcg caacggcagt
     2641 ccttgggaac caacttggca ccgctgtcgg gttcctgctg ccacccgtgc tggtgcctaa
     2701 tacccaaaac gatacgaacc tgctcgcctg caacatttcc actatgttct acggcacctc
     2761 tgcagtggcg actctcctgt tcatactgac cgcgatcgcg ttcaaggaga agccgcgcta
     2821 cccaccgtca caagcccagg ctgccttgca ggattcacca cccgaggaat atagctacaa
     2881 gaagagcatc aggaatttgt tcaagaatat cccgttcgtg ctgttgctga ttacctacgg
     2941 cattatgaca ggggcatttt atagtgtgtc aactctcctc aatcaaatga ttctcacata
     3001 ttatgaaggt gaggaagtga acgcaggcag aatcggcctg acccttgtgg tcgccggcat
     3061 ggtaggtagc atcctgtgcg gtttgtggct cgactacacc aagacgtata agcaaacgac
     3121 ccttattgtc tacatactgt ccttcatcgg catggtcatt ttcaccttta ctctggatct
     3181 gaggtacatc attattgtct tcgtgaccgg tggcgtcctg gggttcttta tgacaggcta
     3241 tctgcccctg gggttcgagt tcgccgtcga gattacatat cccgagtcag aggggacatc
     3301 ttccgggctg ctgaacgcaa gcgcccaaat tttcggtatc ctgtttaccc tggcccaggg
     3361 gaaattgacc tctgattacg gaccgaaagc tggtaatatc ttcctttgcg tgtggatgtt
     3421 cattgggatt atcctgacgg ccctgattaa atccgacctt cggcggcata atatcaacat
     3481 tgggatcacc aacgtcgacg tcaaggccat ccccgccgat agcccgactg atcaggagcc
     3541 taagactgtg atgctgagta aacaatctga gagtgctatc gcgggaggtt caggtggatc
     3601 agactacaaa gaccatgacg gtgattataa agatcatgac attgattaca aggatgacga
     3661 tgacaagtag gctacgtaaa ttccgccccc cccccccctc tccctccccc ccccctaacg
     3721 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3781 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3841 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3901 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     3961 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     4021 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     4081 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     4141 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     4201 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     4261 gatgataata tggccacaac catggttcct ttgtctcaag aagaatccac cctcattgaa
     4321 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     4381 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     4441 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     4501 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     4561 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     4621 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagtc
     4681 gacgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4741 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4801 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4861 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     4921 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4981 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     5041 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     5101 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     5161 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     5221 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     5281 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     5341 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     5401 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5461 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5521 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5581 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5641 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5701 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5761 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5821 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5881 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5941 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6001 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6061 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6121 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6181 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6241 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6301 tcacctagat ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg
     6361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7261 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7321 aataggggtt ccgcgcacat ttcc
//
