LOCUS       sequence-327653-        6999 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 6999)
  TITLE     Gadella lab plasmids
REFERENCE   2  (bases 1 to 6999)
  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"
COMMENT     Sequence Label: pTfR-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2900..2909
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2906..3601
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3039..3057)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3307..3326
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3727..3848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3764..3783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3818..3837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3855..4310)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3992..4013)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4204..4223
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4337..4441
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4443..4800
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4443..4634
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4466..4486)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4651..4786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4835..5629
                     /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             4835..5629
                     /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(4889..4908)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5499..5518
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5817..5836)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5861..5908
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6237..6825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6726..6745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagcg ccaccatgat ggatcaagct agatcagcat tctctaactt
      601 gtttggtgga gaaccattgt catatacccg gttcagcctg gctcggcaag tagatggcga
      661 taacagtcat gtggagatga aacttgctgt agatgaagaa gaaaatgctg acaataacac
      721 aaaggccaat gtcacaaaac caaaaaggtg tagtggaagt atctgctatg ggactattgc
      781 tgtgatcgtc tttttcttga ttggatttat gattggctac ttgggctatt gtaaaggggt
      841 agaaccaaaa actgagtgtg agagactggc aggaaccgag tctccagtga gggaggagcc
      901 aggagaggac ttccctgcag cacgtcgctt atattgggat gacctgaaga gaaagttgtc
      961 ggagaaactg gacagcacag acttcaccag caccatcaag ctgctgaatg aaaattcata
     1021 tgtccctcgt gaggctggat ctcaaaaaga tgaaaatctt gcgttgtatg ttgaaaatca
     1081 atttcgtgaa tttaaactca gcaaagtctg gcgtgatcaa cattttgtta agattcaggt
     1141 caaagacagc gctcaaaact cggtgatcat agttgataag aacggtagac ttgtttacct
     1201 ggtggagaat cctgggggtt atgtggcgta tagtaaggct gcaacagtta ctggtaaact
     1261 ggtccatgct aattttggta ctaaaaaaga ttttgaggat ttatacactc ctgtgaatgg
     1321 atctatagtg attgtcagag cagggaaaat cacctttgca gaaaaggttg caaatgctga
     1381 aagcttaaat gcaattggtg tgttgatata catggaccag actaaatttc ccattgttaa
     1441 cgcagaactt tcattctttg gacatgctca tctggggaca ggtgaccctt acacacctgg
     1501 attcccttcc ttcaaccaca ctcagtttcc accatctcgg tcatcaggat tgcctaatat
     1561 acctgtccag acaatctcca gagctgctgc agaaaagctg tttgggaata tggaaggaga
     1621 ctgtccctct gactggaaaa cagactctac atgtaggatg gtaacctcag aaagcaagaa
     1681 tgtgaagctc actgtgagca atgtgctgaa agagataaaa attcttaaca tctttggagt
     1741 tattaaaggc tttgtagaac cagatcacta tgttgtagtt ggggcccaga gagatgcatg
     1801 gggccctgga gctgcaaaat ccggtgtagg cacagctctc ctattgaaac ttgcccagat
     1861 gttctcagat atggtcttaa aagatgggtt tcagcccagc agaagcatta tctttgccag
     1921 ttggagtgct ggagactttg gatcggttgg tgccactgaa tggctagagg gatacctttc
     1981 gtccctgcat ttaaaggctt tcacttatat taatctggat aaagcggttc ttggtaccag
     2041 caacttcaag gtttctgcca gcccactgtt gtatacgctt attgagaaaa caatgcaaaa
     2101 tgtgaagcat ccggttactg ggcaatttct atatcaggac agcaactggg ccagcaaagt
     2161 tgagaaactc actttagaca atgctgcttt ccctttcctt gcatattctg gaatcccagc
     2221 agtttctttc tgtttttgcg aggacacaga ttatccttat ttgggtacca ccatggacac
     2281 ctataaggaa ctgattgaga ggattcctga gttgaacaaa gtggcacgag cagctgcaga
     2341 ggtcgctggt cagttcgtga ttaaactaac ccatgatgtt gaattgaacc tggactatga
     2401 gaggtacaac agccaactgc tttcatttgt gagggatctg aaccaataca gagcagacat
     2461 aaaggaaatg ggcctgagtt tacagtggct gtattctgct cgtggagact tcttccgtgc
     2521 tacttccaga ctaacaacag atttcgggaa tgctgagaaa acagacagat ttgtcatgaa
     2581 gaaactcaat gatcgtgtca tgagagtgga gtatcacttc ctctctccct acgtatctcc
     2641 aaaagagtct cctttccgac atgtcttctg gggctccggc tctcacacgc tgccagcttt
     2701 actggagaac ttgaaactgc gtaaacaaaa taacggtgct tttaatgaaa cgctgttcag
     2761 aaaccagttg gctctagcta cttggactat tcagggagct gcaaatgccc tctctggtga
     2821 cgtttgggac attgacaatg agttttcaga attcggctcc accggctcca ccggctccac
     2881 cggcgcggat ccaccggtcg ccaccatggt gagcaagggc gaggcagtga tcaaggagtt
     2941 catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg
     3001 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     3061 tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc
     3121 cttcatcaag caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt
     3181 caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac
     3241 ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc
     3301 tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg agcggttgta
     3361 ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg
     3421 ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg
     3481 cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt
     3541 ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     3601 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     3661 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3721 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3781 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3841 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3901 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3961 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4021 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4081 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4141 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4201 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4261 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4321 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4381 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4441 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4501 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4561 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4621 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4681 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4741 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4801 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4861 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4921 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4981 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5041 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5101 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5161 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5221 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5281 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5341 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5401 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5461 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5521 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5581 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5641 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5701 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5761 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5821 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5881 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5941 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6001 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6061 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6121 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6181 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6241 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6301 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6361 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6421 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6481 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6541 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6601 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6661 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6721 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6781 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6841 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6901 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     6961 tacggggtca ttagttcata gcccatatat ggagttccg
//
