LOCUS       sequence-440953-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human FLVCR1b with C-terminal 3X_FLAG cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  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 6516)
  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 FLVCR1b-3X FLAG
FEATURES             Location/Qualifiers
     source          1..6516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2775..2840
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(2868..3442,3443..3445,3446..3454)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3035..3052)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3262..3281
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3455..3850
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4021..4490
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4620..4637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4791..5379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4871..4890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5543..6334,6335..6403))
                     /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     6166..6185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6404..6508)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaatgt
     1921 tttatggaac atcagctgtt gccacacttt tatttatttt aacagcaatt gccttcaaag
     1981 aaaaacctcg gtatccacca agtcaggctc aagcagctct tcaagacagt ccccctgaag
     2041 agtactccta taagaaatca ataagaaacc tgtttaaaaa cattcctttt gtccttctgt
     2101 tgatcactta tggtatcatg actggtgcct tttattcagt ctcaacgtta ttaaatcaaa
     2161 tgatattgac atattatgag ggagaagaag tcaatgctgg aaggattggg ctaacgctag
     2221 tagtagctgg aatggtgggc tctattcttt gtggcttatg gctggattat actaaaacat
     2281 acaaacagac tactctgata gtttatattt tgtcttttat tggaatggtt atctttactt
     2341 tcacattgga ccttagatat attatcatcg tgtttgttac tggaggggtg cttggcttct
     2401 tcatgactgg ttacctccct ttgggttttg aatttgctgt tgaaatcact taccctgaat
     2461 ctgaaggtac ttcatctggt cttcttaatg cttctgcaca gatatttgga attttgttca
     2521 cattggctca aggaaagctc acatcagact atggtcctaa ggcagggaac atttttctct
     2581 gtgtctggat gtttataggc atcatattaa cagcattaat caagtctgat ctgcgaagac
     2641 acaacataaa tataggaatt acaaatgttg atgttaaagc tataccagct gacagtccca
     2701 cagaccaaga accaaaaacg gttatgttgt ccaagcagtc agaatcagca attgcgggag
     2761 gttcaggtgg atcagactac aaagaccatg acggtgatta taaagatcat gacattgatt
     2821 acaaggatga cgatgacaag tgagctacgt aaattccgcc cccccccccc ctctccctcc
     2881 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2941 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3001 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3061 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3121 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3181 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3241 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3301 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3361 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3421 ctttgaaaaa cacgatgata atatggccac aaccatggtt cctttgtctc aagaagaatc
     3481 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     3541 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     3601 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     3661 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     3721 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     3781 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     3841 ggagggctaa gtcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     3901 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     3961 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     4021 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4081 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     4141 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     4201 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     4261 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     4321 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     4381 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     4441 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     4501 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     4561 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4621 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4681 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4741 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4801 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4861 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4921 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4981 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5041 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5101 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5161 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5221 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5281 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5341 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5401 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5461 tcaaaaagga tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat
     5521 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5581 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5641 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5701 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5761 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5821 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5881 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5941 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6001 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6061 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6121 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6181 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6241 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6301 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6361 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6421 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6481 agaaaaataa acaaataggg gttccgcgca catttc
//
