LOCUS       sequence-368436-        6018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eGFP-TM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6018)
  AUTHORS   Shi Z, Graber ZT, Baumgart T, Stone HA, Cohen AE
  TITLE     Cell Membranes Resist Flow.
  JOURNAL   Cell. 2018 Dec 13;175(7):1769-1779.e13. doi: 
            10.1016/j.cell.2018.09.054. Epub 2018 Nov 1.
  PUBMED    30392960
REFERENCE   2  (bases 1 to 6018)
  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"; doi: 
            "10.1016/j.cell.2018.09.054"; journalName: "Cell"; date: 
            "2018-12-13- 13"; volume: "175"; issue: "7"; pages: "1769-1779.e13"
COMMENT     Sequence Label: ZS007:eGFP-TM
FEATURES             Location/Qualifiers
     source          1..6018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(69..88)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        167..358
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     315..335
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(418..1209,1210..1278))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1041..1060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1279..1383)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1410..1865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1497..1516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1707..1728
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        1877..2256
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2257..2460
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2411..2431
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2505..2524
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2505..2523
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     2604..2663
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2667..2693
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2703..2705,2706..3416)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2703..2705,2706..3416)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2745..2766)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3006..3025)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3353..3374
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3435..3464
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3468..3614
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(3636..3653)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3642..3866
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3998..4586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4078..4097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4915..4962
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4987..5006
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5194..5988)
                     /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             complement(5194..5988)
                     /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(5305..5324)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5915..5934
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
       61 aggccgggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
      121 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
      181 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      241 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      301 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
      361 ctcttccttt ttcaataaat caatctaaag tatatatgag taaacttggt ctgacagtta
      421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      541 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      721 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1021 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1261 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1381 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     1441 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1501 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1561 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1621 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1681 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1741 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1801 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     1861 aatttacgcg cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     1921 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     1981 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2041 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2101 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2161 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2221 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2281 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2341 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     2401 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     2461 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
     2521 agggagaccc aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg
     2581 aattcggctt ggggatatcc accatggaga cagacacact cctgctatgg gtactgctgc
     2641 tctgggttcc aggttccact ggtgactatc catatgatgt tccagattat gctggggccc
     2701 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2761 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2821 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2881 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2941 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3001 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3061 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3121 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3181 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3241 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3301 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3361 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagccgc
     3421 ggctgcaggt cgacgaacaa aaactcatct cagaagagga tctgaatgct gtgggccagg
     3481 acacgcagga ggtcatcgtg gtgccacact ccttgccctt taaggtggtg gtgatctcag
     3541 ccatcctggc cctggtggtg ctcaccatca tctcccttat catcctcatc atgctttggc
     3601 agaagaagcc acgttaggcg gccgctcgag atcagcctcg actgtgcctt ctagttgcca
     3661 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3721 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3781 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3841 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagtg gcggtaatac
     3901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4741 aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac
     4801 ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg
     4861 gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa
     4921 acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt
     4981 ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca
     5041 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     5101 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
     5161 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     5221 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     5281 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     5341 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     5401 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgct
     5461 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     5521 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     5581 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     5641 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     5701 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     5761 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca gttcattcag
     5821 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     5881 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     5941 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     6001 tcatcctgtc tcttgatc
//
