LOCUS       sequence-213030-        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        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"
     CDS             complement(1565..1576)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2960..2969
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2966..3661
                     /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(3099..3117)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3367..3386
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3787..3908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3824..3843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3878..3897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3915..4370)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4052..4073)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4264..4283
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4397..4501
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4503..4860
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4503..4694
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4526..4546)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4711..4846
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4895..5689
                     /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             4895..5689
                     /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(4949..4968)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5559..5578
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5877..5896)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5921..5968
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6297..6885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6786..6805
                     /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 ttcaatcaca 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 gaggcagtga tcaaggagtt
     3001 catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg
     3061 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     3121 tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc
     3181 cttcatcaag caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt
     3241 caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac
     3301 ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc
     3361 tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcatccaccg agcggttgta
     3421 ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg
     3481 ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg
     3541 cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt
     3601 ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     3661 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     3721 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3781 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3841 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3901 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3961 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4021 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4081 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4141 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4201 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4261 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4321 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4381 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4441 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4501 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4561 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4621 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4681 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4741 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4801 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4861 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4921 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4981 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5041 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5101 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5161 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5221 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5281 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5341 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5401 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5461 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5521 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5581 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5641 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5701 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5761 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5821 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5881 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5941 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6001 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6061 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6121 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6181 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6241 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6301 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6361 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6421 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6481 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6541 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6601 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6661 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6721 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6781 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6841 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6901 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6961 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
