LOCUS       sequence-216919-        5348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transmembrane model receptor to probe receptor internalization via 
            clathrin mediated pathway.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5348)
  AUTHORS   DeGroot ACM, Busch DJ, Hayden CC, Mihelic SA, Alpar AT, Behar M, 
            Stachowiak JC
  TITLE     Entropic Control of Receptor Recycling Using Engineered Ligands.
  JOURNAL   Biophys J. 2018 Mar 27;114(6):1377-1388. doi: 
            10.1016/j.bpj.2018.01.036.
  PUBMED    29590595
REFERENCE   2  (bases 1 to 5348)
  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.bpj.2018.01"; journalName: "Biophys J"; date: 
            "2018-03-27- 27"; volume: "114"; issue: "6"; pages: "1377-1388"
COMMENT     Sequence Label: TfR-Delta-EctoR1-mRFP-GFPnb
FEATURES             Location/Qualifiers
     source          1..5348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        126..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             985..1659
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             985..1659
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1115..1133)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1383..1402
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1577..1600
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1714..2055
                     /codon_start=1
                     /product="GFP-binding fragment of a single-chain camelid 
                     antibody (Rothbauer et al., 2008)"
                     /label=GFP nanobody
                     /translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
                     WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCNVNVGFEYW
                     GQGTQVTVSS"
     CDS             2059..2076
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2201..2322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2238..2257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2292..2311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2329..2784)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2466..2487)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2678..2697
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2811..2915
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2917..3274
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2917..3108
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2940..2960)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3125..3260
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3309..4103
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3309..4103
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3363..3382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3973..3992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4291..4310)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4335..4382
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4711..5299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5200..5219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
       61 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      661 cgctaccgga ctcagatctc gagctcaagc ttcgaattca tgatggatca agctagatca
      721 gcattctcta acttgtttgg tggagaacca ttgtcatata cccggttcag cctggctcgg
      781 caagtagatg gcgataacag tcatgtggag atgaaacttg ctgtagatga agaagaaaat
      841 gctgacaata acacaaaggc caatgtcaca aaaccaaaaa ggtgtagtgg aagtatctgc
      901 tatgggacta ttgctgtgat cgtctttttc ttgattggat ttatgattgg ctacttgggc
      961 tatggtaaag gggatccacc ggttatggcc tcctccgagg acgtcatcaa ggagttcatg
     1021 cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1081 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     1141 cccctgccct tcgcctggga catcctgtcc cctcagttcc agtacggctc caaggcctac
     1201 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     1261 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     1321 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     1381 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg gatgtacccc
     1441 gaggacggcg ccctgaaggg cgagatcaag atgaggctga agctgaagga cggcggccac
     1501 tacgacgccg aggtcaagac cacctacatg gccaagaagc ccgtgcagct gcccggcgcc
     1561 tacaagaccg acatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     1621 cagtacgagc gcgccgaggg ccgccactcc accggcgccg aatcgcggcc gcatgtgcgg
     1681 ccgcaataca atcaaaggag atataccatg caggttcaac tggtggaaag cggcggtgct
     1741 ctggtacaac cgggcggtag tctgcgcctg agctgtgccg caagcggttt cccagtcaac
     1801 cgctactcta tgcgttggta tcgccaggcg cctggtaaag aacgtgaatg ggttgccggc
     1861 atgagcagtg cgggcgatcg ttctagttac gaggactctg ttaaaggtcg ttttacaatt
     1921 agccgtgatg atgcgcgcaa taccgtgtat ctgcaaatga acagtctgaa gccggaggac
     1981 accgcagtat attattgcaa tgtcaacgtg gggtttgaat attggggcca ggggactcag
     2041 gtgacggtga gctctaaaca tcaccatcac catcactaag cggccgcgac tctagatcat
     2101 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2161 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2281 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2341 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2401 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2461 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2521 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2581 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2641 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2701 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2761 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2821 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2881 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2941 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3001 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3061 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3121 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3181 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3241 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3301 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3481 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4081 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4141 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4201 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4261 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4321 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4381 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4441 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4501 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4561 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4621 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5341 ctggcctt
//
