LOCUS       sequence-350020-        6586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of Folate Receptor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6586)
  AUTHORS   Chen C, Ke J, Zhou XE, Yi W, Brunzelle JS, Li J, Yong EL, Xu HE, 
            Melcher K
  TITLE     Structural basis for molecular recognition of folic acid by folate 
            receptors.
  JOURNAL   Nature. 2013 Aug 22;500(7463):486-9. doi: 10.1038/nature12327. Epub 
            2013 Jul 14.
  PUBMED    23851396
REFERENCE   2  (bases 1 to 6586)
  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: 
            "10.1038/nature12327"; journalName: "Nature"; date: "2013-08-22- 
            22"; volume: "500"; issue: "7463"; pages: "486-9"
COMMENT     Sequence Label: pCDNA6-IgK-hFRa(23-234)C11S-th-Fc
FEATURES             Location/Qualifiers
     source          1..6586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..95)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        80..409
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      260..395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     322..341
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        457..504
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             523..921
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1079..1200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1116..1135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1170..1189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1249..1265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1249..1265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1262..1284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1273..1289
                     /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(1297..1303,1304..1321,1322..1327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1342..1363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1480..1497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1651..2239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1731..1750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2410..3201,3202..3270))
                     /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     3033..3052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3271..3375)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3450..3469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3641..4020
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4021..4224
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4175..4195
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4269..4288
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4269..4287
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4307..4316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(4313..4357,4440..4451)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGVHS"
     CDS             join(4313..4358,4441..4451)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGVHS"
     primer_bind     4423..4442
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5100..5117
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5814..5831
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5857..5898
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5908..5925
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5948..5965)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5954..6178
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6224..6586,1..66)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6311..6330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6521..6542
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
       61 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
      121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
      181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
      241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
      361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
      421 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
      481 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gcctttgtct
      541 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
      601 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
      661 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
      721 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
      781 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
      841 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
      901 ggttatgtgt gggagggcta agcacttcgt ggccgaggag caggactgac acgtgctacg
      961 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     1021 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     1081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1201 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     1261 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1321 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1381 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1441 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1501 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1561 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1621 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2161 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2341 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2401 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2461 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2521 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2581 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2641 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2701 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2761 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2821 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2881 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2941 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3001 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3061 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3121 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3181 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3241 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3301 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3361 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     3421 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3481 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     3541 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     3601 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     3661 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     3721 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     3781 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     3841 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     3901 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     3961 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4021 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     4081 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     4141 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     4201 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
     4261 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgcca ccatgggatg
     4321 gagctgtatc atcctcttct tggtagcaac agctacaggt aaggggctca cagtagcagg
     4381 cttgaggtct ggacatatat atgggtgaca atgacatcca ctttgccttt ctctccacag
     4441 gtgtccactc cgaattccag acaaggattg catgggccag gactgagctt ctcaatgtct
     4501 gcatgaacgc caagcaccac aaggaaaagc caggccccga ggacaagttg catgagcagt
     4561 gtcgaccctg gaggaagaat gcctgctgtt ctaccaacac cagccaggaa gcccataagg
     4621 atgtttccta cctatataga ttcaactgga accactgtgg agagatggca cctgcctgca
     4681 aacggcattt catccaggac acctgcctct acgagtgctc ccccaacttg gggccctgga
     4741 tccagcaggt ggatcagagc tggcgcaaag agcgggtact gaacgtgccc ctgtgcaaag
     4801 aggactgtga gcaatggtgg gaagattgtc gcacctccta cacctgcaag agcaactggc
     4861 acaagggctg gaactggact tcagggttta acaagtgcgc agtgggagct gcctgccaac
     4921 ctttccattt ctacttcccc acacccactg ttctgtgcaa tgaaatctgg actcactcct
     4981 acaaggtcag caactacagc cgagggagtg gccgctgcat ccagatgtgg ttcgacccag
     5041 cccagggcaa ccccaatgag gaggtggcga ggttctatgc tgcagccatg agtggtaccc
     5101 tggttccgcg tggttccgag cccaaatcta gtgacaaaac tcacacatgc ccaccgtgcc
     5161 cagcacctga actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca
     5221 ccctcatgat ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag
     5281 accctgaggt caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa
     5341 agccgcggga ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc
     5401 accaggactg gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag
     5461 cccccatcga gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca
     5521 ccctgccccc atcccgggat gagctgacca agaaccaggt cagcctgacc tgcctggtca
     5581 aaggcttcta tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca
     5641 actacaaggc cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc
     5701 tcaccgtgga caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg
     5761 aggctctgca caaccactac acgcagaaga gcctctccct gtctccgggt aaacatcacc
     5821 accatcacca ttgataattc tagagggccc ttcgaaggta agcctatccc taaccctctc
     5881 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc
     5941 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6001 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6061 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6121 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     6181 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     6241 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6301 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6361 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6421 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6481 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6541 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataag
//
