LOCUS       sequence-261583-        7011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of Transferrin receptors (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7011)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 7011)
  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: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: pTfR-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..7011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2960..2969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2966..3676
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3221..3230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3799..3920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3836..3855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3890..3909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3927..4382)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4064..4085)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4276..4295
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4409..4513
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4515..4872
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4515..4706
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4538..4558)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4723..4858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4907..5701
                     /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             4907..5701
                     /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(4961..4980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5571..5590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5889..5908)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5933..5980
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6309..6897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6798..6817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcg ccaccatgat ggatcaagct agatcagcat tctctaactt
      661 gtttggtgga gaaccattgt catatacccg gttcagcctg gctcggcaag tagatggcga
      721 taacagtcat gtggagatga aacttgctgt agatgaagaa gaaaatgctg acaataacac
      781 aaaggccaat gtcacaaaac caaaaaggtg tagtggaagt atctgctatg ggactattgc
      841 tgtgatcgtc tttttcttga ttggatttat gattggctac ttgggctatt gtaaaggggt
      901 agaaccaaaa actgagtgtg agagactggc aggaaccgag tctccagtga gggaggagcc
      961 aggagaggac ttccctgcag cacgtcgctt atattgggat gacctgaaga gaaagttgtc
     1021 ggagaaactg gacagcacag acttcaccag caccatcaag ctgctgaatg aaaattcata
     1081 tgtccctcgt gaggctggat ctcaaaaaga tgaaaatctt gcgttgtatg ttgaaaatca
     1141 atttcgtgaa tttaaactca gcaaagtctg gcgtgatcaa cattttgtta agattcaggt
     1201 caaagacagc gctcaaaact cggtgatcat agttgataag aacggtagac ttgtttacct
     1261 ggtggagaat cctgggggtt atgtggcgta tagtaaggct gcaacagtta ctggtaaact
     1321 ggtccatgct aattttggta ctaaaaaaga ttttgaggat ttatacactc ctgtgaatgg
     1381 atctatagtg attgtcagag cagggaaaat cacctttgca gaaaaggttg caaatgctga
     1441 aagcttaaat gcaattggtg tgttgatata catggaccag actaaatttc ccattgttaa
     1501 cgcagaactt tcattctttg gacatgctca tctggggaca ggtgaccctt acacacctgg
     1561 attcccttcc ttcaaccaca ctcagtttcc accatctcgg tcatcaggat tgcctaatat
     1621 acctgtccag acaatctcca gagctgctgc agaaaagctg tttgggaata tggaaggaga
     1681 ctgtccctct gactggaaaa cagactctac atgtaggatg gtaacctcag aaagcaagaa
     1741 tgtgaagctc actgtgagca atgtgctgaa agagataaaa attcttaaca tctttggagt
     1801 tattaaaggc tttgtagaac cagatcacta tgttgtagtt ggggcccaga gagatgcatg
     1861 gggccctgga gctgcaaaat ccggtgtagg cacagctctc ctattgaaac ttgcccagat
     1921 gttctcagat atggtcttaa aagatgggtt tcagcccagc agaagcatta tctttgccag
     1981 ttggagtgct ggagactttg gatcggttgg tgccactgaa tggctagagg gatacctttc
     2041 gtccctgcat ttaaaggctt tcacttatat taatctggat aaagcggttc ttggtaccag
     2101 caacttcaag gtttctgcca gcccactgtt gtatacgctt attgagaaaa caatgcaaaa
     2161 tgtgaagcat ccggttactg ggcaatttct atatcaggac agcaactggg ccagcaaagt
     2221 tgagaaactc actttagaca atgctgcttt ccctttcctt gcatattctg gaatcccagc
     2281 agtttctttc tgtttttgcg aggacacaga ttatccttat ttgggtacca ccatggacac
     2341 ctataaggaa ctgattgaga ggattcctga gttgaacaaa gtggcacgag cagctgcaga
     2401 ggtcgctggt cagttcgtga ttaaactaac ccatgatgtt gaattgaacc tggactatga
     2461 gaggtacaac agccaactgc tttcatttgt gagggatctg aaccaataca gagcagacat
     2521 aaaggaaatg ggcctgagtt tacagtggct gtattctgct cgtggagact tcttccgtgc
     2581 tacttccaga ctaacaacag atttcgggaa tgctgagaaa acagacagat ttgtcatgaa
     2641 gaaactcaat gatcgtgtca tgagagtgga gtatcacttc ctctctccct acgtatctcc
     2701 aaaagagtct cctttccgac atgtcttctg gggctccggc tctcacacgc tgccagcttt
     2761 actggagaac ttgaaactgc gtaaacaaaa taacggtgct tttaatgaaa cgctgttcag
     2821 aaaccagttg gctctagcta cttggactat tcagggagct gcaaatgccc tctctggtga
     2881 cgtttgggac attgacaatg agttttcaga attcggctcc accggctcca ccggctccac
     2941 cggcgcggat ccaccggtcg ccaccatggt gagcaagggc gaggaggata acatggcctc
     3001 tctcccagcg acacatgagt tacacatctt tggctccatc aacggtgtgg actttgacat
     3061 ggtgggtcag ggcaccggca atccaaatga tggttatgag gagttaaacc tgaagtccac
     3121 caagggtgac ctccagttct ccccctggat tctggtccct catatcgggt atggcttcca
     3181 tcagtacctg ccctaccctg acgggatgtc gcctttccag gccgccatgg tagatggctc
     3241 cggctaccaa gtccatcgca caatgcagtt tgaagatggt gcctccctta ctgttaacta
     3301 ccgctacacc tacgagggaa gccacatcaa aggagaggcc caggtgaagg ggactggttt
     3361 ccctgctgac ggtcctgtga tgaccaactc gctgaccgct gcggactggt gcaggtcgaa
     3421 gaagacttac cccaacgaca aaaccatcat cagtaccttt aagtggagtt acaccactgg
     3481 aaatggcaag cgctaccgga gcactgcgcg gaccacctac acctttgcca agccaatggc
     3541 ggctaactat ctgaagaacc agccgatgta cgtgttccgt aagacggagc tcaagcactc
     3601 caagaccgag ctcaacttca aggagtggca aaaggccttt accgatgtga tgggcatgga
     3661 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     3721 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3781 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3841 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3901 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3961 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4021 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4081 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4141 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4201 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4261 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4321 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4381 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4441 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4501 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4561 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4621 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4681 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4741 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4801 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4861 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4921 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4981 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5041 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     5101 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5161 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5221 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5281 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5341 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5401 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     5461 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5521 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5581 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5641 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5701 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5761 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5821 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5881 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5941 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     6001 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     6061 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     6121 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     6181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
