LOCUS       sequence-446223-        5495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Hisx6-Tagged msYFP in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5495)
  TITLE     Yellow and oxidation-resistant derivatives of a monomeric 
            superfolder GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5495)
  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            "Unpublished"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: pQE81XN-msYFP
FEATURES             Location/Qualifiers
     source          1..5495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      160..169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(208..229)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     816..837
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      952..1046
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1090..1749
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1156..1175)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1814..1900
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1993..3075)
                     /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"
     primer_bind     3037..3056
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3076..3153)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3237..3259)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3345..3485
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3400..3419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3500..3517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3671..4259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3751..3770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4430..5221,5222..5290))
                     /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     5053..5072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5291..5395)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5463..5481
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatctcacc atcaccatca ccatacggat ccactagtcg ccaccatgga tagcactgag
      181 agcctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      241 aagttcagcg tgcgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag
      301 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
      361 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      421 tccgccatgc ccgaaggcta cgtccaggag cgcaccatca ccttcaagga cgacggcacc
      481 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      541 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc
      601 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
      661 aagatccgcc acaacgtgga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
      721 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
      781 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      841 gccgccggga tcactcacgg cagctccggc tccagcgggt aaagcggccg cgactctaga
      901 tcactcggta ccccgggtcg acctgcagcc aagcttaatt agctgagctt ggactcctgt
      961 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1021 gccgggcgtt ttttattggt gagaatccaa gctagcttgg cgagattttc aggagctaag
     1081 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1141 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1201 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1261 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     1321 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1381 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1441 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1501 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     1561 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcactatggg caaatattat
     1621 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1681 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1741 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc
     1801 tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     1861 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccctcta gattacgtgc
     1921 agtcgatgat aagctgtcaa acatgagaat tgtgcctaat gagtgagcta acttacatta
     1981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     2101 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     2161 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     2221 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     2281 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     2341 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     2401 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     2461 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     2521 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     2581 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     2641 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     2701 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     2761 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     2821 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     2881 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     2941 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     3001 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     3061 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     3121 accgcgaaag gttttgcacc attcgatggt gtcggaattt cgggcagcgt tgggtcctgg
     3181 ccacgggtgc gcatgatcta gagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3241 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3301 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     3361 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     3421 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3481 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3541 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3601 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3661 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3721 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3781 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3841 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3901 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3961 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4021 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4081 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4141 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4201 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4261 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4321 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4381 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4441 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4501 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4561 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4621 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4681 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4741 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4801 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4861 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4921 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4981 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5041 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5101 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5161 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5221 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5281 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5341 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5401 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5461 aaaataggcg tatcacgagg ccctttcgtc ttcac
//
