LOCUS       sequence-377008-        4332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express 6xHis-Tag miniGFP2 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4332)
  AUTHORS   Liang GT, Lai C, Yue Z, Zhang H, Li D, Chen Z, Lu X, Tao L, Subach 
            FV, Piatkevich KD
  TITLE     Enhanced small green fluorescent proteins as a multisensing platform
            for biosensor development.
  JOURNAL   Front Bioeng Biotechnol. 2022 Oct 17;10:1039317. doi: 
            10.3389/fbioe.2022.1039317. eCollection 2022.
  PUBMED    36324888
REFERENCE   2  (bases 1 to 4332)
  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: "Front 
            Bioeng Biotechnol."; date: "2022-10-17"; volume: "10:1039317. doi"; 
            pages: " 10.3389/fbioe.2022.1039317. eCollection 2022"
COMMENT     Sequence Label: pBAD-HisD-miniGFP2
FEATURES             Location/Qualifiers
     source          1..4332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             328..345
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(733..750)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      903..989
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1081..1108
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1127..1218
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1219..1287,1288..2079)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPXRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1437..1456)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2250..2838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2739..2758
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2992..3009
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3024..3164
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3090..3109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3250..3272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3428..4306)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggagccac catcaccatc accatggcag atctatggag
      361 aagtcctttg tgattacaga cccatggctg cccgactacc ctattatttc cgctagtgac
      421 ggattcctgg agctgaccga gtattctcgg gatgaaatca tgggaaggaa cgcacgcttc
      481 ctgcagggac ccgagactga ccaggccacc gtgcagaaga tccgggacgc tattcgagat
      541 cggagaccta ccacagtcca gctgatcaac tacacaaaga gcggcaagaa gttctggaat
      601 ctgttgcacc tgcagcctgt gtttgacgga aaaggcgggc tgcagtattt tattggagtg
      661 cagctggtcg gctccgatca tgtctaagaa ttcgaagctt ggctgttttg gtggatgaga
      721 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
      781 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
      841 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
      901 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      961 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     1021 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     1081 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     1141 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1201 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1261 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1321 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1381 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1441 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1501 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1561 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1621 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1681 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1741 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     1801 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1861 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1921 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagcccycc
     1981 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2041 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2101 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2161 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2221 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2281 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2341 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2401 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2461 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2521 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2581 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2641 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2701 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2761 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2821 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2881 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2941 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3001 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3061 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3121 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3181 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3241 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3301 ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg
     3361 acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg
     3421 ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     3481 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     3541 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
     3601 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     3661 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     3721 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     3781 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     3841 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     3901 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     3961 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     4021 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     4081 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     4141 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     4201 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     4261 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     4321 cgccattcag ag
//
