LOCUS       sequence-291877-        6329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP-SHB fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6329)
  AUTHORS   Pietila I, Van Mourik D, Tamelander A, Kriz V, Claesson-Welsh L, 
            Tengholm A, Welsh M
  TITLE     Temporal Dynamics of VEGFA-Induced VEGFR2/FAK Co-Localization Depend
            on SHB.
  JOURNAL   Cells. 2019 Dec 15;8(12). pii: cells8121645. doi: 
            10.3390/cells8121645.
  PUBMED    31847469
REFERENCE   2  (bases 1 to 6329)
  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: "Cells."; 
            date: "2019-12-15"; volume: "8(12). pii: cells8121645. doi"; pages: 
            " 10.3390/cells8121645"
COMMENT     Sequence Label: peGFP-N1-SHB
FEATURES             Location/Qualifiers
     source          1..6329
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3117..3238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3154..3173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3208..3227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3245..3700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3382..3403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3594..3613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3727..3831
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3833..4190
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3833..4024
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3856..3876)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4041..4176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4225..5019
                     /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             4225..5019
                     /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(4279..4298)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4889..4908
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5207..5226)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5251..5298
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5627..6215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6116..6135
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccatggccaa gtggctaaac aagtacttca gcttgggcaa cagcaagacc
     1381 aagagccccc cgcagccgcc gcggccagac taccgcgagc agcggcgccg aggcgagcgg
     1441 ccttcgcagc ccccccaggc cgtgccgcag gcctcctccg ccgcctcggc gtcctgcggt
     1501 ccggccaccg cctcctgctt ctcagcctct tcgggctcgc tgcccgacga cagcggcagc
     1561 accagcgacc tcatccgcgc ctaccgcgcg cagaaggagc gagacttcga ggacccctac
     1621 aacgggcctg gctcgtcgct gcgcaaactg cgcgccatgt gccgcctgga ctactgcggc
     1681 ggcagcgggg agccaggcgg ggtccagcgc gccttctcgg cctcgtccgc gtcgggcgcc
     1741 gcgggctgtt gctgcgcctc ctcgggcgcg ggggccgccg cgtcctcgtc ctcgtcctcc
     1801 ggctctccgc atctctaccg cagcagcagc gagcggcggc ccgccacgcc ggccgaggtg
     1861 cgctacatct cccccaagca ccgcctcatc aaagtggaga gcgccgcggg cggtggggcc
     1921 ggggaccccc tggggggcgc ctgcgcgggc ggccgcacct ggagcccgac ggcctgcgga
     1981 ggcaagaaac tgctcaacaa gtgcgccgcc tcagccgcgg aggagagcgg ggccggcaag
     2041 aaggacaagg tgaccatagc cgatgactac tcagatccct ttgatgccaa gaatgatctc
     2101 aagagcaaag caggaaaggg ggagagtgct ggctacatgg agccctatga ggcacagagg
     2161 atcatgacag aatttcagag gcaggaaagt gtccggtccc agcataaagg tatccagtta
     2221 tatgacaccc cttacgaacc tgaaggccaa agtgttgact cagactcgga gagcacagtc
     2281 agcccccgac tgcgggagag caagctgccc caggatgacg acaggcccgc cgatgagtac
     2341 gaccagcctt gggagtggaa ccgggtcacc atcccagccc tggcagcaca gtttaatggc
     2401 aacgagaagc ggcagtcatc cccctcacct tcgcgggacc ggcggcgcca gcttcgtgcc
     2461 cctggagggg gctttaagcc tatcaaacat gggagccctg agttctgcgg gatcctagga
     2521 gaaagggtgg atcctgccgt ccccctggag aagcaaatat ggtatcacgg agccatcagc
     2581 agaggagacg ccgagaacct gctgcgactc tgcaaggagt gtagctacct tgtccggaac
     2641 agccagacca gcaagcatga ctactccctc tccctgagga gcaaccaggg ttttatgcac
     2701 atgaaactgg ccaaaaccaa agagaaatac gttctgggtc agaacagccc tccgttcgac
     2761 agtgtcccgg aagtcatcca ctactacacc accagaaagc tacccatcaa aggggctgag
     2821 cacttgtccc tcctctatcc cgtggctgtg aggaccctgt gagcggacca gacctgccct
     2881 gctctgtgac agagcctgag acttggaggt gccagaggcc ccccaccaac cagcccagcc
     2941 actgttgctg gctgtgtcgt ttgtgttgtg tgtatggtac tagcacacca ctgcatgtct
     3001 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3061 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     3241 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3361 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3421 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3481 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3541 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3601 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3661 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3721 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3781 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3841 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4021 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4081 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4141 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4201 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4261 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4321 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4381 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4441 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4501 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4561 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4621 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4681 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4741 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4801 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4861 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4921 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4981 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5161 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5221 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5281 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5341 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5401 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5461 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5521 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5761 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6301 ctgtggataa ccgtattacc gccatgcat
//
