LOCUS       sequence-233932-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mGinger1 tagged N-terminally with an MHHHHHHGSG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  AUTHORS   Wannier TM, Gillespie SK, Hutchins N, McIsaac RS, Wu SY, Shen Y, 
            Campbell RE, Brown KS, Mayo SL
  TITLE     Monomerization of far-red fluorescent proteins.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Nov 27;115(48):E11294-E11301. doi: 
            10.1073/pnas.1807449115. Epub 2018 Nov 13.
  PUBMED    30425172
REFERENCE   2  (bases 1 to 5807)
  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: 
            "10.1073/pnas.1807449115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-11-27- 27"; volume: "115"; issue: "48"; pages: 
            "E11294-E11301"
COMMENT     Sequence Label: pET-mGinger1
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     812..831
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(851..928)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1134..1153
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1241..1260
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1241..1259
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1260..1284
                     /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             1299..1321
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1331..1348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2045..2068
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2095..2113)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2109..2156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2193..2648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2280..2299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2490..2511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2763..3554,3555..3623))
                     /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     3386..3405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3624..3716)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3797..4385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4286..4305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4539..4556
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4571..4710
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4636..4655)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4796..4818
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4812..5003)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5575..5807,1..850))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
       61 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
      121 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
      181 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
      241 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
      301 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
      361 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
      421 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
      481 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
      541 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
      601 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
      661 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
      721 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
      781 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
      841 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
      901 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
      961 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     1021 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     1081 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     1141 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     1201 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg
     1261 gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag
     1321 atataccatg caccaccatc atcaccatgg cagcggcctg ctgaaagaaa atatgcatat
     1381 caagatgtat atggaaggca ccgtgaatgg tcactacttt aagtgtaagg gtgaaggcga
     1441 tggtaatcca ttcgctggca cacagtccat gcgtgttcat gttaccgagg gcgctccgct
     1501 ccctttcgcc tttgacatcc tcgccccgtg ctgtgaatat ggctctaaga cctttgttcg
     1561 ctatccagcg gatatcccgg actttttcaa acagtccttc ccggagggct tcacttggga
     1621 acgtacgact acctacgagg acggtggtat cctgaccgcg caccaggaca ccagcctgga
     1681 gggtaactgc ctgatttaca aggttaaggt tcatggcacg aatttcccgg ctgacggtcc
     1741 ggttatgaaa aaggaaactt gtggttggga accgtccacc gagagcgttt acccggaaaa
     1801 cggtgttttg cgcggtcgta atcacatggc gctcaaggtt ggtgatagcc acctccactg
     1861 ccaccactcc acgacctacc gctctaaaaa ggcggaacgc gccctgatta tgccgccgcg
     1921 ccacagcact gattactgtc tgcaaataac gagtcataag gatgatgagt acttcgagct
     1981 gtacgaaacg tctgtggcgc gttactctga ctaatgagct ttcttgtaca aagtggtttc
     2041 cgcttggagc cacccgcagt tcgaaaagta agtgattaac ctaggctgct gccaccgctg
     2101 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2161 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     2221 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2281 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2341 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2401 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2461 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2521 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2581 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2641 tacaatttct ggcggcacga tggcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3421 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3481 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3541 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3601 acacggaaat gttgaatact catactcttc ctttttcaat catgattgaa gcatttatca
     3661 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3781 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3901 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3961 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4021 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4081 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4141 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4201 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4261 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4321 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4381 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4441 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4501 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4561 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4621 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4681 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4741 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4801 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4861 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4921 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4981 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     5041 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     5101 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     5161 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     5221 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     5281 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     5341 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     5401 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     5461 ctagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     5521 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     5581 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5641 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     5701 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     5761 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcg
//
