LOCUS       sequence-271349-        7019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Transferrin Receptor, Excitation: 485, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7019)
  TITLE     Davidson Green fluorescent proteins
REFERENCE   2  (bases 1 to 7019)
  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: sfGFP-TFR-20
FEATURES             Location/Qualifiers
     source          1..7019
                     /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             join(2906..2908,2909..2911,2912..3625)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2951..2972)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3559..3580
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3747..3868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3784..3803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3838..3857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3875..4330)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4012..4033)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4224..4243
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4357..4461
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4463..4820
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4463..4654
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4486..4506)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4671..4806
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4855..5649
                     /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             4855..5649
                     /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(4909..4928)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5519..5538
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5837..5856)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5881..5928
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6257..6845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6746..6765
                     /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 gaggagctgt tcaccggggt
     2941 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg
     3001 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     3061 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     3121 cagccgctac cccgaccaca tgaagcgcca cgacttcttc aagtccgcca tgcccgaagg
     3181 ctacgtccag gagcgcacca tcagcttcaa ggacgacggc acctacaaga cccgcgccga
     3241 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3301 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta
     3361 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
     3421 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3481 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgtgctga gcaaagaccc
     3541 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
     3601 cggcatggac gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca
     3661 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3721 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3781 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3841 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3901 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3961 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4021 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4081 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4141 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4201 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4261 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4321 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4381 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4441 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4501 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4621 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4681 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4741 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4801 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4861 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4921 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4981 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5041 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5101 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5161 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5221 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5281 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5341 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5401 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5461 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5521 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5581 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5641 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5701 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5761 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5821 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5881 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5941 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6001 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6061 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6901 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     6961 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
