LOCUS       sequence-324214-        7013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Transferin Receptor, Excitation: 399, Emission: 456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7013)
  AUTHORS   Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV
  TITLE     An enhanced monomeric blue fluorescent protein with the high 
            chemical stability of the chromophore.
  JOURNAL   PLoS One. 2011;6(12):e28674. Epub 2011 Dec 8.
  PUBMED    22174863
REFERENCE   2  (bases 1 to 7013)
  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: "PLoS One.";
            date: "2011"; volume: "6(12)"; pages: "e28674. Epub 2011 Dec 8"
COMMENT     Sequence Label: mTagBFP2-TFR-20
FEATURES             Location/Qualifiers
     source          1..7013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        111..414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..618
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3010..3019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3016..3726
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    3851..3972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3888..3907)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3942..3961
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3979..4434)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4116..4137)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4328..4347
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4461..4565
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4567..4924
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4567..4758
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4590..4610)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4775..4910
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4959..5753
                     /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             4959..5753
                     /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(5013..5032)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5623..5642
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5941..5960)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5985..6032
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6361..6949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6850..6869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
       61 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      121 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      181 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      241 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      301 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      361 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      601 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag
      661 atctcgagcg ccaccatgat ggatcaagct agatcagcat tctctaactt gtttggtgga
      721 gaaccattgt catatacccg gttcagcctg gctcggcaag tagatggcga taacagtcat
      781 gtggagatga aacttgctgt agatgaagaa gaaaatgctg acaataacac aaaggccaat
      841 gtcacaaaac caaaaaggtg tagtggaagt atctgctatg ggactattgc tgtgatcgtc
      901 tttttcttga ttggatttat gattggctac ttgggctatt gtaaaggggt agaaccaaaa
      961 actgagtgtg agagactggc aggaaccgag tctccagtga gggaggagcc aggagaggac
     1021 ttccctgcag cacgtcgctt atattgggat gacctgaaga gaaagttgtc ggagaaactg
     1081 gacagcacag acttcaccag caccatcaag ctgctgaatg aaaattcata tgtccctcgt
     1141 gaggctggat ctcaaaaaga tgaaaatctt gcgttgtatg ttgaaaatca atttcgtgaa
     1201 tttaaactca gcaaagtctg gcgtgatcaa cattttgtta agattcaggt caaagacagc
     1261 gctcaaaact cggtgatcat agttgataag aacggtagac ttgtttacct ggtggagaat
     1321 cctgggggtt atgtggcgta tagtaaggct gcaacagtta ctggtaaact ggtccatgct
     1381 aattttggta ctaaaaaaga ttttgaggat ttatacactc ctgtgaatgg atctatagtg
     1441 attgtcagag cagggaaaat cacctttgca gaaaaggttg caaatgctga aagcttaaat
     1501 gcaattggtg tgttgatata catggaccag actaaatttc ccattgttaa cgcagaactt
     1561 tcattctttg gacatgctca tctggggaca ggtgaccctt acacacctgg attcccttcc
     1621 ttcaaccaca ctcagtttcc accatctcgg tcatcaggat tgcctaatat acctgtccag
     1681 acaatctcca gagctgctgc agaaaagctg tttgggaata tggaaggaga ctgtccctct
     1741 gactggaaaa cagactctac atgtaggatg gtaacctcag aaagcaagaa tgtgaagctc
     1801 actgtgagca atgtgctgaa agagataaaa attcttaaca tctttggagt tattaaaggc
     1861 tttgtagaac cagatcacta tgttgtagtt ggggcccaga gagatgcatg gggccctgga
     1921 gctgcaaaat ccggtgtagg cacagctctc ctattgaaac ttgcccagat gttctcagat
     1981 atggtcttaa aagatgggtt tcagcccagc agaagcatta tctttgccag ttggagtgct
     2041 ggagactttg gatcggttgg tgccactgaa tggctagagg gatacctttc gtccctgcat
     2101 ttaaaggctt tcacttatat taatctggat aaagcggttc ttggtaccag caacttcaag
     2161 gtttctgcca gcccactgtt gtatacgctt attgagaaaa caatgcaaaa tgtgaagcat
     2221 ccggttactg ggcaatttct atatcaggac agcaactggg ccagcaaagt tgagaaactc
     2281 actttagaca atgctgcttt ccctttcctt gcatattctg gaatcccagc agtttctttc
     2341 tgtttttgcg aggacacaga ttatccttat ttgggtacca ccatggacac ctataaggaa
     2401 ctgattgaga ggattcctga gttgaacaaa gtggcacgag cagctgcaga ggtcgctggt
     2461 cagttcgtga ttaaactaac ccatgatgtt gaattgaacc tggactatga gaggtacaac
     2521 agccaactgc tttcatttgt gagggatctg aaccaataca gagcagacat aaaggaaatg
     2581 ggcctgagtt tacagtggct gtattctgct cgtggagact tcttccgtgc tacttccaga
     2641 ctaacaacag atttcgggaa tgctgagaaa acagacagat ttgtcatgaa gaaactcaat
     2701 gatcgtgtca tgagagtgga gtatcacttc ctctctccct acgtatctcc aaaagagtct
     2761 cctttccgac atgtcttctg gggctccggc tctcacacgc tgccagcttt actggagaac
     2821 ttgaaactgc gtaaacaaaa taacggtgct tttaatgaaa cgctgttcag aaaccagttg
     2881 gctctagcta cttggactat tcagggagct gcaaatgccc tctctggtga cgtttgggac
     2941 attgacaatg agttttcaga attcggctcc accggctcca ccggctccac cggcgcggat
     3001 ccaccggtcg ccaccatggt gtctaagggc gaagagctga ttaaggagaa catgcacatg
     3061 aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa
     3121 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
     3181 cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac
     3241 cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag
     3301 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     3361 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct
     3421 gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac
     3481 ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc
     3541 gcaaacgcca agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc
     3601 gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc
     3661 gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag
     3721 cttaattaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3781 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3841 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3901 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3961 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4021 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4081 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4141 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4201 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4261 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4321 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4381 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4441 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4561 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4621 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4681 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4741 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4801 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4861 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4921 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4981 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5041 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5101 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5161 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5221 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5281 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5341 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5401 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5461 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5521 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5581 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5641 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5701 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5761 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5821 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5881 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5941 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6001 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6061 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6121 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6181 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6241 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6301 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6361 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6421 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6481 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6541 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6601 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6661 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6721 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6781 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6841 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6901 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6961 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tct
//
