LOCUS       sequence-223018-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP.Strands 1-9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  AUTHORS   Caldwell RM, Bermudez JG, Thai D, Aonbangkhen C, Schuster BS, 
            Courtney T, Deiters A, Hammer DA, Chenoweth DM, Good MC
  TITLE     Optochemical Control of Protein Localization and Activity within 
            Cell-like Compartments.
  JOURNAL   Biochemistry. 2018 May 8;57(18):2590-2596. doi: 
            10.1021/acs.biochem.8b00131. Epub 2018 Apr 19.
  PUBMED    29671583
REFERENCE   2  (bases 1 to 4937)
  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/acs.biochem.8b00131"; journalName: "Biochemistry"; date: 
            "2018-05-8- 8"; volume: "57"; issue: "18"; pages: "2590-2596"
COMMENT     Sequence Label: pRC01
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1204)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="TKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHAD
                     ELYRSCLARFGDDGENL"
     primer_bind     1716..1735
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1716..1734
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1766..1788
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1811..2461
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     primer_bind     2422..2444
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2498..2518
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3118..3135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3153..3170
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3223..3241)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3237..3284
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3321..3776
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3408..3427)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3618..3639
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3802..3906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3907..3975,3976..4767)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4125..4144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac tagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggttactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     1321 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     1381 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     1441 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     1501 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     1561 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     1621 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     1681 acccaacgct gcccgagatc tcgatcccgc gaaattaata cgactcacta tagggagacc
     1741 acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgaa
     1801 acaagatctg atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     1861 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     1921 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     1981 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     2041 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     2101 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     2161 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     2221 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     2281 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     2341 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     2401 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     2461 aactagtgga tctggtggtg gtggcggttc catgagcgag aatctttatt ttcagggatc
     2521 catgcgcaaa ggagaagaac ttttcactgg agttgtccca attcttattg aattagatgg
     2581 tgatgttaat gggcacaaat ttttcgtccg tggagagggt gaaggtgatg ctacaatcgg
     2641 aaaactcagc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     2701 tgtcactact ctgacctatg gtgttcaatg cttttcccgt tatccggatc acatgaaacg
     2761 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatactt
     2821 caaagatgac gggacctaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt
     2881 taatcgtatc gagttaaaag gtattgattt taaagaagat ggaaacattc tcggacacaa
     2941 actcgagtac aactttaact cacacaaggt atacatcacg gcagacaaac aaaacaatgg
     3001 aatcaaagct aacttcacca ttcgccacaa cgttgaagat ggttccgttc aactagcaga
     3061 ccattatcaa caaaatactc caattggcga tggccctgtc tgatagtaag cggccgccac
     3121 catcaccacc accactaggg gccgcactcg agcaccacca ccaccaccac tgagatccgg
     3181 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     3241 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     3301 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3361 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3421 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3481 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3541 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3601 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3661 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3721 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     3781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3901 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3961 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4021 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4081 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4141 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4201 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4261 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4321 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4381 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4441 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4501 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4561 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4621 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4681 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4741 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4921 aagatcaaag gatcttc
//
