LOCUS       sequence-350692-        4135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of His-tagged roGFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4135)
  TITLE     pQE-60 roGFP2-His
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4135)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pQE-60 roGFP2-His
FEATURES             Location/Qualifiers
     source          1..4135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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)."
     protein_bind    53..69
                     /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)."
     RBS             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(106..108,109..111,112..822)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(151..172)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(412..431)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     759..780
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             829..846
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      869..963
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1073..1092)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1735..1821
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1868..1890)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1976..2116
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2031..2050
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2131..2148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2302..2890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2382..2401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3061..3852,3853..3921))
                     /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     3684..3703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3922..4026)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4094..4112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taaccatggt gagcaagggc
      121 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      181 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      241 aagttcatct ccaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      301 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      361 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      421 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      481 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      541 tacaactgcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      601 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      661 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacctgc
      721 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      781 accgccgccg ggatcactct cggcatggac gagctgtaca agagatctca tcaccatcac
      841 catcactaag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      901 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      961 aatccaagct agcttggcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     1021 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1081 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     1141 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     1201 cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg
     1261 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     1321 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     1381 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     1441 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     1501 cttcttcgcc cccgttttca ccatgcatgg gcaaatatta tacgcaaggc gacaaggtgc
     1561 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     1621 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     1681 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
     1741 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1801 gctctcctga gtaggacaaa tccgccctct agagctgcct cgcgcgtttc ggtgatgacg
     1861 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     1921 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     1981 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     2041 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2101 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2701 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     2761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2821 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2881 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2941 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3001 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3061 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3121 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3181 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3241 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3301 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3361 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3421 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3481 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3541 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3601 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3661 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3721 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3781 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3841 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3901 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3961 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4021 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4081 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg agaaa
//
