LOCUS       sequence-319423-        9279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human TFRC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9279)
  AUTHORS   Frank M, Citarella CG, Quinones GB, Bentley M
  TITLE     A Novel Labeling Strategy Reveals That Myosin Va and Myosin Vb Bind 
            the Same Dendritically Polarized Vesicle Population.
  JOURNAL   Traffic. 2020 Sep 21. doi: 10.1111/tra.12764.
  PUBMED    32959500
REFERENCE   2  (bases 1 to 9279)
  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: "Traffic. 
            2020 Sep 21. doi: 10.1111/tra.12764."
COMMENT     Sequence Label: pBa.TfR-Halo
FEATURES             Location/Qualifiers
     source          1..9279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..276
                     /label=chicken beta-actin promoter
     primer_bind     1219..1241
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             complement(2250..2261)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3606..3615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3636..3656
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3672..4556
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    4698..4832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4735..4754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4789..4808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4863..4879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4863..4879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4876..4898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4887..4903
                     /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(4911..4917,4918..4935,4936..4941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4956..4977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5094..5111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5265..5853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5345..5364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6024..6815,6816..6884))
                     /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     6647..6666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6885..6989)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7057..7075
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7113..7135)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7235..7254
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7448..7470
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7462..7479
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7463..7479
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
       61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      121 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      181 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      241 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      301 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      361 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      421 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
      481 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
      541 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
      601 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
      661 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
      721 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc
      781 acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg
      841 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
      901 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
      961 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1021 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     1081 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     1141 gccgtcccct tctccatctc cagcctcggg gctgtccgca gggggacggc tgccttcggg
     1201 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggggtt tatatcttcc
     1261 cttctctgtt cctccgcagc ccccaagctt ggcgcgccta ggggggatcc accatggatc
     1321 aagctagatc agcattctct aacttgtttg gtggagaacc attgtcatat acccggttca
     1381 gcctggctcg gcaagtagat ggcgataaca gtcatgtgga gatgaaactt gctgtagatg
     1441 aagaagaaaa tgctgacaat aacacaaagg ccaatgtcac aaaaccaaaa aggtgtagtg
     1501 gaagtatctg ctatgggact attgctgtga tcgtcttttt cttgattgga tttatgattg
     1561 gctacttggg ctattgtaaa ggggtagaac caaaaactga gtgtgagaga ctggcaggaa
     1621 ccgagtctcc agtgagggag gagccaggag aggacttccc tgcagcacgt cgcttatatt
     1681 gggatgacct gaagagaaag ttgtcggaga aactggacag cacagacttc accagcacca
     1741 tcaagctgct gaatgaaaat tcatatgtcc ctcgtgaggc tggatctcaa aaagatgaaa
     1801 atcttgcgtt gtatgttgaa aatcaatttc gtgaatttaa actcagcaaa gtctggcgtg
     1861 atcaacattt tgttaagatt caggtcaaag acagcgctca aaactcggtg atcatagttg
     1921 ataagaacgg tagacttgtt tacctggtgg agaatcctgg gggttatgtg gcgtatagta
     1981 aggctgcaac agttactggt aaactggtcc atgctaattt tggtactaaa aaagattttg
     2041 aggatttata cactcctgtg aatggatcta tagtgattgt cagagcaggg aaaatcacct
     2101 ttgcagaaaa ggttgcaaat gctgaaagct taaatgcaat tggtgtgttg atatacatgg
     2161 accagactaa atttcccatt gttaacgcag aactttcatt ctttggacat gctcatctgg
     2221 ggacaggtga cccttacaca cctggattcc cttccttcaa tcacactcag tttccaccat
     2281 ctcggtcatc aggattgcct aatatacctg tccagacaat ctccagagct gctgcagaaa
     2341 agctgtttgg gaatatggaa ggagactgtc cctctgactg gaaaacagac tctacatgta
     2401 ggatggtaac ctcagaaagc aagaatgtga agctcactgt gagcaatgtg ctgaaagaga
     2461 taaaaattct taacatcttt ggagttatta aaggctttgt agaaccagat cactatgttg
     2521 tagttggggc ccagagagat gcatggggcc ctggagctgc aaaatccggt gtaggcacag
     2581 ctctcctatt gaaacttgcc cagatgttct cagatatggt cttaaaagat gggtttcagc
     2641 ccagcagaag cattatcttt gccagttgga gtgctggaga ctttggatcg gttggtgcca
     2701 ctgaatggct agagggatac ctttcgtccc tgcatttaaa ggctttcact tatattaatc
     2761 tggataaagc ggttcttggt accagcaact tcaaggtttc tgccagccca ctgttgtata
     2821 cgcttattga gaaaacaatg caaaatgtga agcatccggt tactgggcaa tttctatatc
     2881 aggacagcaa ctgggccagc aaagttgaga aactcacttt agacaatgct gctttccctt
     2941 tccttgcata ttctggaatc ccagcagttt ctttctgttt ttgcgaggac acagattatc
     3001 cttatttggg taccaccatg gacacctata aggaactgat tgagaggatt cctgagttga
     3061 acaaagtggc acgagcagct gcagaggtcg ctggtcagtt cgtgattaaa ctaacccatg
     3121 atgttgaatt gaacctggac tatgagaggt acaacagcca actgctttca tttgtgaggg
     3181 atctgaacca atacagagca gacataaagg aaatgggcct gagtttacag tggctgtatt
     3241 ctgctcgtgg agacttcttc cgtgctactt ccagactaac aacagatttc gggaatgctg
     3301 agaaaacaga cagatttgtc atgaagaaac tcaatgatcg tgtcatgaga gtggagtatc
     3361 acttcctctc tccctacgta tctccaaaag agtctccttt ccgacatgtc ttctggggct
     3421 ccggctctca cacgctgcca gctttactgg agaacttgaa actgcgtaaa caaaataacg
     3481 gtgcttttaa tgaaacgctg ttcagaaacc agttggctct agctacttgg actattcagg
     3541 gagctgcaaa tgccctctct ggtgacgttt gggacattga caatgagttt ggggatccac
     3601 cggtcgccac catggctagc ctcgagccaa ccactgagga tctgtacttt cagagcgata
     3661 acgatggatc cgaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg
     3721 gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc
     3781 acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc
     3841 atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt
     3901 atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag
     3961 aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca
     4021 atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg
     4081 acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc
     4141 gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc
     4201 gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
     4261 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
     4321 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
     4381 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
     4441 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
     4501 cggacctgat cggcagcgag atcgcgcgct ggctgtctac tctggagatt tccggtctgt
     4561 acaagcggac cgtctaatct agataagtaa tgatcataat cagccatatc acatctgtag
     4621 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4681 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4741 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4801 aactcatcaa tgtatcttat catgtctgga tccccgggta ccgagctcga attcgtaatc
     4861 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7021 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7081 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     7141 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     7201 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     7261 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     7321 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7381 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7441 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagttggg atctttgcat
     7501 tggcccacgg ctctcaggat ggggatgctc cccttcagca cccggttccc cttggaaact
     7561 gatggtcctg gctctgtggc atggcagtgg cactgtgagg agccccctac cagcagcaca
     7621 cagtggggtt ggcactgcca cgctccggat gccgcgctct gatccaaccc cataatcaag
     7681 ggaacccgaa ttgccccatc attgccccca ccacccccat cctgccgggc cctcacaccc
     7741 cacgctgcct tctggtgaca ttccccagcc caaacccacg gcttcatggc taccgcgggg
     7801 catttcccat tgccgcccca ttatcagctc tgcacacctc ccgctgtacc catgcctcgt
     7861 ggctgccctt ctttgacgta taatcttcta attaataccc ggccttgtca aagtggagca
     7921 caaacgttaa ttaattcccc agcaggcagg taattaacag tgtgactccc tttttgctgc
     7981 gagtggggct gatacagaga gatgtggcac tatggagccc acggggtcct ggcactgggt
     8041 gcccacggag gtccccatgt gctgcagtgt caccgcctcc gaggagacag tattgtccct
     8101 gcggtgtccc tgcagctcag ctctgtccac agggccacct ccagtttgga ggggacacaa
     8161 tgcagccccg atgcaaccca tcctcgcagc atcccaggga caaagacccc actgcaagac
     8221 cgcacacagg gctgggtccc gctcccctaa tatctacagt gcttcttgca tggcccctta
     8281 atcaatgcag ttaatcagca tgcgctcatg caccgctctg gagctgcaaa gcccctcgca
     8341 gcgctgctca ccaacaccgc gcaccgcccc ggcccagcct gcagcacgcg ctgcaaacag
     8401 gaaagaaaca aaatattgcc caaatgtagg caaaggcatt cggctgcctt gacctccgcc
     8461 gggccgggcc ctgcctgact cagctcctta ctcagcgctc gcttcctccc tccggctgcc
     8521 accgccgcag cgcacaccct gacaaagagt ggcccttaac gggctctgag gtgcacccag
     8581 cagtgcactc agcagtccaa gggccggcct ggaggtttgc accgctacgt gctgacatta
     8641 gcattgaact tggccctggg tagtgctgca ggccgggcgg ggtgggtgta gagagtgcag
     8701 cgcgcgttgc acccggtgcc ccttcccctc ccttgcatcc cagcaggctg caccccagca
     8761 ccaggcccgt gcatgcatgc tcctggtgtt attgcagcct ggtgcatgca tgcgtcttga
     8821 tggtgcagcg ctgtgcatgc atcctccttg gtgtgtagca gtctagtgca tgcatacccc
     8881 tcggtgttat tgctgctctg tgcacgcacg ctcattgtat cacttcatcc cagtgcatgc
     8941 actcacactg gagcgattgc tgctcggtgc acgcacactc attgtatcac tgcagctcag
     9001 tggctgcacg cacaccggtg ttattgctgc tcggtgcgtg catgcacatc agtgtcgctg
     9061 cagctcagtg catgcacgct cattgcccat cgctatccct gcctctcctg ctggcgctcc
     9121 ccgggaggtg acttcaaggg gaccgcagga ccacctcggg ggtgggggga gggctgcaca
     9181 cgcggacccc gctccccctc cccaacaaag cactgtggaa tcaaaaaggg gggagggggg
     9241 atggaggggc gcgtcacacc cccgccccac accctcacc
//
