LOCUS       sequence-287372-        7773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human transferrin receptor with an HA tag fused at the C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7773)
  AUTHORS   Smyth JW, Vogan JM, Buch PJ, Zhang SS, Fong TS, Hong TT, Shaw RM
  TITLE     Actin cytoskeleton rest stops regulate anterograde traffic of 
            connexin 43 vesicles to the plasma membrane.
  JOURNAL   Circ Res. 2012 Mar 30;110(7):978-89. Epub 2012 Feb 9.
  PUBMED    22328533
REFERENCE   2  (bases 1 to 7773)
  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: "Circ Res.";
            date: "2012-03-30"; volume: "110(7)"; pages: "978-89. Epub 2012 Feb 
            9"
COMMENT     Sequence Label: pcDNA3.2/DEST/hTfR-HA
FEATURES             Location/Qualifiers
     source          1..7773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    913..937
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1882..1893)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3227..3251)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3268..3294
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3346..3365)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3390..3438
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3640..4068
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3727..3746)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3937..3958
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4077..4097)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4082..4411
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4262..4397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4324..4343
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4478..5272
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4532..4551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5142..5161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5448..5569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5485..5504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5539..5558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5618..5634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5618..5634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5631..5653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5642..5658
                     /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(5666..5672,5673..5690,5691..5696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5711..5732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5849..5866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6020..6608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6100..6119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6779..7570,7571..7639))
                     /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     7402..7421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7640..7744)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gttaagctat caacaagttt gtacaaaaaa gcaggctcca ccatgatgga tcaagctaga
      961 tcagcattct ctaacttgtt tggtggagaa ccattgtcat atacccggtt cagcctggct
     1021 cggcaagtag atggcgataa cagtcatgtg gagatgaaac ttgctgtaga tgaagaagaa
     1081 aatgctgaca ataacacaaa ggccaatgtc acaaaaccaa aaaggtgtag tggaagtatc
     1141 tgctatggga ctattgctgt gatcgtcttt ttcttgattg gatttatgat tggctacttg
     1201 ggctattgta aaggggtaga accaaaaact gagtgtgaga gactggcagg aaccgagtct
     1261 ccagtgaggg aggagccagg agaggacttc cctgcagcac gtcgcttata ttgggatgac
     1321 ctgaagagaa agttgtcgga gaaactggac agcacagact tcaccggcac catcaagctg
     1381 ctgaatgaaa attcatatgt ccctcgtgag gctggatctc aaaaagatga aaatcttgcg
     1441 ttgtatgttg aaaatcaatt tcgtgaattt aaactcagca aagtctggcg tgatcaacat
     1501 tttgttaaga ttcaggtcaa agacagcgct caaaactcgg tgatcatagt tgataagaac
     1561 ggtagacttg tttacctggt ggagaatcct gggggttatg tggcgtatag taaggctgca
     1621 acagttactg gtaaactggt ccatgctaat tttggtacta aaaaagattt tgaggattta
     1681 tacactcctg tgaatggatc tatagtgatt gtcagagcag ggaaaatcac ctttgcagaa
     1741 aaggttgcaa atgctgaaag cttaaatgca attggtgtgt tgatatacat ggaccagact
     1801 aaatttccca ttgttaacgc agaactttca ttctttggac atgctcatct ggggacaggt
     1861 gacccttaca cacctggatt cccttccttc aatcacactc agtttccacc atctcggtca
     1921 tcaggattgc ctaatatacc tgtccagaca atctccagag ctgctgcaga aaagctgttt
     1981 gggaatatgg aaggagactg tccctctgac tggaaaacag actctacatg taggatggta
     2041 acctcagaaa gcaagaatgt gaagctcact gtgagcaatg tgctgaaaga gataaaaatt
     2101 cttaacatct ttggagttat taaaggcttt gtagaaccag atcactatgt tgtagttggg
     2161 gcccagagag atgcatgggg ccctggagct gcaaaatccg gtgtaggcac agctctccta
     2221 ttgaaacttg cccagatgtt ctcagatatg gtcttaaaag atgggtttca gcccagcaga
     2281 agcattatct ttgccagttg gagtgctgga gactttggat cggttggtgc cactgaatgg
     2341 ctagagggat acctttcgtc cctgcattta aaggctttca cttatattaa tctggataaa
     2401 gcggttcttg gtaccagcaa cttcaaggtt tctgccagcc cactgttgta tacgcttatt
     2461 gagaaaacaa tgcaaaatgt gaagcatccg gttactgggc aatttctata tcaggacagc
     2521 aactgggcca gcaaagttga gaaactcact ttagacaatg ctgctttccc tttccttgca
     2581 tattctggaa tcccagcagt ttctttctgt ttttgcgagg acacagatta tccttatttg
     2641 ggtaccacca tggacaccta taaggaactg attgagagga ttcctgagtt gaacaaagtg
     2701 gcacgagcag ctgcagaggt cgctggtcag ttcgtgatta aactaaccca tgatgttgaa
     2761 ttgaacctgg actatgagag gtacaacagc caactgcttt catttgtgag ggatctgaac
     2821 caatacagag cagacataaa ggaaatgggc ctgagtttac agtggctgta ttctgctcgt
     2881 ggagacttct tccgtgctac ttccagacta acaacagatt tcgggaatgc tgagaaaaca
     2941 gacagatttg tcatgaagaa actcaatgat cgtgtcatga gagtggagta tcacttcctc
     3001 tctccctacg tatctccaaa agagtctcct ttccgacatg tcttctgggg ctccggctct
     3061 cacacgctgc cagctttact ggagaacttg aaactgcgta aacaaaataa cggtgctttt
     3121 aatgaaacgc tgttcagaaa ccagttggct ctagctactt ggactattca gggagctgca
     3181 aatgccctct ctggtgacgt ttgggacatt gacaatgagt ttttggaccc agctttcttg
     3241 tacaaagtgg ttgatctagg cggcccgtac ccatacgacg tcccagatta cgcgctcgag
     3301 tctacgcgta ccggttagta atgagtttaa acgggggagg ctaactgaaa cacggaagga
     3361 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3421 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     3481 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     3541 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     3601 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3661 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3721 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3781 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3841 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3901 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3961 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4021 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4081 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4141 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4201 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4261 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4321 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4381 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4441 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4501 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4561 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4621 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4681 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4741 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4801 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4861 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4921 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4981 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5041 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5101 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5161 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5221 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5281 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5341 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5401 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5461 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5521 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5581 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5641 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5701 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5761 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5821 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5881 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5941 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6001 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6061 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6121 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6181 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6241 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6301 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6361 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6421 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6481 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6541 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgagc agaaaaaaag
     6601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7381 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7741 cgcgcacatt tccccgaaaa gtgccacctg acg
//
