LOCUS       sequence-383724-        4752 bp    DNA     circular SYN 15-APR-2025
DEFINITION  titratable expression of mEos3.2 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4752)
  AUTHORS   Shilling PJ, Khananisho D, Cumming AJ, Soderstrom B, Daley DO
  TITLE     Signal amplification of araC pBAD using a standardized translation 
            initiation region.
  JOURNAL   Synth Biol (Oxf). 2022 Jul 5;7(1):ysac009. doi: 
            10.1093/synbio/ysac009. eCollection 2022.
  PUBMED    35903559
REFERENCE   2  (bases 1 to 4752)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synth Biol 
            (Oxf)."; date: "2022-07-5"; volume: "7(1):ysac009. doi"; pages: " 
            10.1093/synbio/ysac009. eCollection 2022"
COMMENT     Sequence Label: pBAD/HisB (TIRSTD)-mEos3.2
FEATURES             Location/Qualifiers
     source          1..4752
                     /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             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /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(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             430..1107
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="SAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNARN
                     DITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     primer_bind     complement(1153..1170)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1323..1409
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1501..1528
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1547..1638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1639..1707,1708..2499)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1857..1876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2670..3258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3159..3178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3412..3429
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3444..3584
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3510..3529)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3670..3692
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3848..4726)
                     /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 ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagatcta gtgcgattaa gccagacatg aagatcaaac tccgtatgga aggcaacgta
      481 aacgggcacc actttgtgat cgacggagat ggtacaggca agccttttga gggaaaacag
      541 agtatggatc ttgaagtcaa agagggcgga cctctgcctt ttgcctttga tatcctgacc
      601 actgcattcc attacggcaa cagggtattc gccaaatatc cagacaacat acaagactat
      661 tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa gcttgacttt cgaagacggg
      721 ggcatttgca atgccagaaa cgacataaca atggaagggg acactttcta taataaagtt
      781 cgattttatg gtaccaactt tcccgccaat ggtccagtta tgcagaagaa gacgctgaaa
      841 tgggagccct ccactgagaa aatgtatgtg cgtgatggag tgctgacggg tgatattgaa
      901 atggctttgt tgcttgaagg aaatgcccat taccgatgtg acttcagaac tacttacaaa
      961 gctaaggaga agggtgtcaa gttaccaggc gcacactttg tggaccactg cattgagatt
     1021 ttaagccatg acaaagatta caacaaggtt aagctgtatg agcatgctgt tgctcattct
     1081 ggattgcctg acaatgccag acgataagaa ttcgaagctt ggctgttttg gcggatgaga
     1141 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     1201 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     1261 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     1321 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     1381 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     1441 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     1501 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     1561 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1621 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1681 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1741 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1801 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1861 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1921 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1981 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2041 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2101 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2161 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2221 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2281 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2341 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2401 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2461 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2521 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2581 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2821 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2881 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2941 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3001 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3061 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3121 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3181 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3241 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3301 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3361 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3421 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3481 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3541 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3601 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3661 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3721 ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg
     3781 acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg
     3841 ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     3901 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     3961 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtagtgctc aaaagcagct
     4021 tcgcctgact aatgcgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     4081 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     4141 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     4201 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     4261 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     4321 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     4381 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     4441 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     4501 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     4561 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     4621 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     4681 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     4741 cgccattcag ag
//
