LOCUS       sequence-216383-        3901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP.Strand10-Haloenzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3901)
  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 3901)
  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: pRC02
FEATURES             Location/Qualifiers
     source          1..3901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..141
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             145..177
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(181..189,190..204)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             343..1233
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(1302..1320)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1316..1363
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1460..1915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1547..1566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1757..1778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1941..2045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2046..2114,2115..2906)
                     /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     complement(2264..2283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3077..3665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3566..3585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3819..3836
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatc atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg
      181 gatctgtacg acgatgacga taaggatcga tggggatcct acacgatgga cctgccagac
      241 gaccattacc tgtcgacaca aactatcctt tcgaaagatt tgaacggagg cggggggtca
      301 gaaggtggtg gtagtggcgg ggcggccgca ggttcgggtt cgatggcaga aatcggtact
      361 ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat
      421 gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac
      481 gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg
      541 atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc
      601 ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac
      661 tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt
      721 gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc
      781 gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac
      841 gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg
      901 gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca
      961 aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg
     1021 gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg
     1081 atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac
     1141 atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc
     1201 gcgcgctggc tgtcgacgct cgagatttcc ggttaataag aattcgaagc ttgatccggc
     1261 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1321 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1381 atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1441 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1501 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1561 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1621 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1681 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1741 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1801 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1861 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     1921 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1981 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2041 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2101 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2161 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2221 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2281 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2341 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2401 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2461 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2521 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2581 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2641 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2701 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2761 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2821 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2881 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3841 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3901 g
//
