LOCUS       sequence-117808-        7556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP Fluorescent Elastin-like Polypeptide(VPGVG) with 96 repeats 
            containing Valine guest residues.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7556)
  AUTHORS   Pastuszka MK, Janib SM, Weitzhandler I, Okamoto CT, Hamm-Alvarez S, 
            Mackay JA
  TITLE     A tunable and reversible platform for the intracellular formation of
            genetically engineered protein microdomains.
  JOURNAL   Biomacromolecules. 2012 Nov 12;13(11):3439-44. doi: 
            10.1021/bm301090x. Epub 2012 Oct 25.
  PUBMED    23088632
REFERENCE   2  (bases 1 to 7556)
  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: "10.1021/bm301090x";
            journalName: "Biomacromolecules"; date: "2012-11-12- 12"; volume: 
            "13"; issue: "11"; pages: "3439-44"
COMMENT     Sequence Label: GFP-V96 ELP
FEATURES             Location/Qualifiers
     source          1..7556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(904..906,907..909,910..1620)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(937..965)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1560..1580
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3146..3163)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3152..3376
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3422..3850
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3509..3528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3719..3740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3859..3879)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3864..4194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4045..4180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4107..4126
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4261..5055
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4315..4334)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4925..4944
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5229..5350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5266..5285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5320..5339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5399..5415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5399..5415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5412..5434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5423..5439
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5447..5453,5454..5471,5472..5477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5492..5513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5630..5647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5801..6389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5881..5900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6560..7351,7352..7420))
                     /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     7183..7202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7421..7525)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagac
      901 accatggcca gcaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
      961 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgctaca
     1021 tacggaaagc ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
     1081 acacttgtca ctactttctc ttatggtgtt caatgctttt cccgttatcc ggatcatatg
     1141 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata
     1201 tctttcaaag atgacgggaa ctacaagacg cgtgctgaag tcaagtttga aggtgatacc
     1261 cttgttaatc gtatcgagtt aaaaggtatt gattttaaag aagatggaaa cattctcgga
     1321 cacaaactcg agtacaacta taactcacac aatgtataca tcacggcaga caaacaaaag
     1381 aatggaatca aagctaactt caaaattcgc cacaacattg aagatggatc cgttcaacta
     1441 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     1501 cattacctgt cgacacaatc tgccctttcg aaagatccca acgaaaagcg tgaccacatg
     1561 gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga gctctacaaa
     1621 agcggntccg gaccggtgct agcgataggt gtaccaggtg tcggtgtacc gggtgtcggc
     1681 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     1741 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     1801 gttccgggtg taggtgttcc gggcgtgggt gtaccaggtg tcggtgtacc gggtgtcggc
     1861 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     1921 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     1981 gttccgggtg taggtgttcc gggcgtgggt gtaccaggtg tcggtgtacc gggtgtcggc
     2041 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     2101 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     2161 gttccgggtg taggtgttcc gggcgtgggt gtaccaggtg tcggtgtacc gggtgtcggc
     2221 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     2281 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     2341 gttccgggtg taggtgttcc gggcgtgggt gtaccaggtg tcggtgtacc gggtgtcggc
     2401 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     2461 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     2521 gttccgggtg taggtgttcc gggcgtgggt gtaccaggtg tcggtgtacc gggtgtcggc
     2581 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     2641 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     2701 gttccgggtg taggtgttcc gggcgtgggt gtaccaggtg tcggtgtacc gggtgtcggc
     2761 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     2821 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     2881 gttccgggtg taggtgttcc gggcgtgggt gtaccaggtg tcggtgtacc gggtgtcggc
     2941 gtacctggcg tcggtgtccc gggtgttggt gttccgggtg taggtgttcc gggcgtgggt
     3001 gtaccaggtg tcggtgtacc gggtgtcggc gtacctggcg tcggtgtccc gggtgttggt
     3061 gttccgggtg taggtgttcc gggcgtgggt tactgataat gatcttcagg atccgaattc
     3121 gagctccgtc aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3181 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3241 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3301 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3361 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3421 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3481 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3541 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     3601 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3661 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3721 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3781 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3841 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3901 caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3961 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4021 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4081 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4141 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4201 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4261 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4321 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4381 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4441 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4501 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4561 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4621 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4681 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4741 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4801 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4861 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4921 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4981 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5041 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5101 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5161 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5221 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5281 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5341 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5401 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5461 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5521 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5581 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5641 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5701 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5761 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5821 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5881 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5941 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     6001 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6061 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6241 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7441 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7501 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
