LOCUS       sequence-46660-e        4710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4710)
  AUTHORS   Shcherbakova DM, Hink MA, Joosen L, Gadella TW, Verkhusha VV
  TITLE     An Orange Fluorescent Protein with a Large Stokes Shift for 
            Single-Excitation Multicolor FCCS and FRET Imaging.
  JOURNAL   J Am Chem Soc. 2012 May 9;134(18):7913-23. doi: 10.1021/ja3018972
  PUBMED    22486524
REFERENCE   2  (bases 1 to 4710)
  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"; journalName: 
            "J Am Chem Soc"; date: "2012-05-9- 9"; volume: "134"; issue: "18"; 
            pages: "7913-23"
COMMENT     Sequence Label: pBAD-LSSmOrange
FEATURES             Location/Qualifiers
     source          1..4710
                     /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"
     CDS             355..1065
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     768..787
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1111..1128)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1281..1367
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1459..1486
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1505..1596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1597..1665,1666..2457)
                     /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(1815..1834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2628..3216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3117..3136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3370..3387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3402..3542
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3468..3487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3628..3650
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3806..4684)
                     /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 atctatggtg
      361 agcaagggcg aggagaataa catggccatc atcaaggagt tcatgcgctt caaggtgcgc
      421 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
      481 tacgagggct ttcagaccgt taagctgaag gtgaccaagg gtggccccct gcccttcgcc
      541 tgggacatct tgtcccctca gttcacctac ggctccaagg cctacgtgaa gcaccccgcc
      601 gacatccccg actacctcaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
      661 aacttcgagg acggcggcgt ggtgaccgtg actcaggact cctccctgca ggacggcgag
      721 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
      781 aagaagacca tgggcatgga ggcctcctcc gagcggatgt accccgagga cggcgccctg
      841 aagggcgagg acaagctcag gctgaagctg aaggacggcg gccactacac ctccgaggtc
      901 aagaccacct acaaggccaa gaagcccgtg cagttgcccg gcgcctacat cgtcgacatc
      961 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     1021 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagaatt cgaagcttgg
     1081 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     1141 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     1201 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     1261 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1321 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     1381 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     1441 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     1501 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1561 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1621 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1681 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1741 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1801 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     1861 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1921 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1981 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2041 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2101 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2161 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2221 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2281 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2341 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2401 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2461 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2521 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2581 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2641 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2701 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2761 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2821 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2881 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2941 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3001 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3061 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3121 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3181 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3241 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3301 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3361 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3421 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     3481 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3541 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3601 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3661 catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat
     3721 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     3781 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     3841 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     3901 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     3961 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     4021 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     4081 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     4141 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     4201 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     4261 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     4321 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     4381 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     4441 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     4501 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     4561 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     4621 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     4681 ccatactttt catactcccg ccattcagag
//
