LOCUS       sequence-223490-        7020 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 7020)
  TITLE     Gadella lab plasmids
REFERENCE   2  (bases 1 to 7020)
  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-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..7020
                     /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             join(2966..2968,2969..2971,2972..3685)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3011..3032)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3272..3291)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3619..3640
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3808..3929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3845..3864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3899..3918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3936..4391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4073..4094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4285..4304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4418..4522
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4524..4881
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4524..4715
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4547..4567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4732..4867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4916..5710
                     /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             4916..5710
                     /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(4970..4989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5580..5599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5898..5917)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5942..5989
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6318..6906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6807..6826
                     /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 gaggagctgt tcaccggggt
     3001 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3061 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3121 caagctgccc gtgccctggc ccaccctcgt gaccaccctg tcctggggcg tgcagtgctt
     3181 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3241 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3301 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3361 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tactttagcg acaacgtcta
     3421 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     3481 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3541 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     3601 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     3661 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     3721 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3781 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3841 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3901 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3961 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4021 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4081 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4141 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4201 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4261 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4321 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4381 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4441 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4501 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4561 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4621 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4681 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4741 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4801 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4861 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4921 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4981 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5041 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5101 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5161 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5221 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5281 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5341 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5401 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5461 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5521 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5581 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5641 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5701 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5761 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5821 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5881 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5941 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6001 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     6061 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6121 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6181 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6541 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6601 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6661 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6721 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6781 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6841 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6901 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6961 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
