LOCUS       sequence-38133-e        4776 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 4776)
  AUTHORS   Gunewardene MS, Subach FV, Gould TJ, Penoncello GP, Gudheti MV, 
            Verkhusha VV, Hess ST
  TITLE     Superresolution imaging of multiple fluorescent proteins with highly
            overlapping emission spectra in living cells.
  JOURNAL   Biophys J. 2011 Sep 21;101(6):1522-8. Epub 2011 Sep 20.
  PUBMED    21943434
REFERENCE   2  (bases 1 to 4776)
  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: "Biophys 
            J."; date: "2011-09-21"; volume: "101(6)"; pages: "1522-8. Epub 2011
            Sep 20"
COMMENT     Sequence Label: pBAD/HisB-PAmKate
FEATURES             Location/Qualifiers
     source          1..4776
                     /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..1131
                     /codon_start=1
                     /product="far-red monomeric fluorescent protein with an 
                     emission peak at 657 nm (Morozova et al., 2010)"
                     /label=TagRFP657
                     /note="derived from mKate"
                     /translation="MSELITENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRGDMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1177..1194)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1347..1433
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1525..1552
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1571..1662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1663..1731,1732..2523)
                     /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(1881..1900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2694..3282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3183..3202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3436..3453
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3468..3608
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3534..3553)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3694..3716
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3872..4750)
                     /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 tgagcgagct gatcaccgag aacatgcaca tgaagctgta catggagggc
      481 accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
      541 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
      601 ctggctacca gcttcatgta cggcagcaag accttcatca accacaccca gggcatcccc
      661 gacttctgga agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
      721 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
      781 aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca
      841 ctcggctggg aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga
      901 ggggacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt gaagaccaca
      961 tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccgc
     1021 agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct
     1081 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatta agaattcgaa
     1141 gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     1201 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     1261 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     1321 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1381 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     1441 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     1501 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     1561 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1621 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1681 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1741 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1801 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1861 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     1921 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1981 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2041 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2101 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2161 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2221 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2281 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2341 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2401 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2461 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2521 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2581 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2641 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2701 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2761 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2821 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2881 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2941 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3001 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3061 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3121 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3181 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3241 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3301 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3361 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3421 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3481 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     3541 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3601 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3661 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3721 caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc
     3781 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg
     3841 tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca
     3901 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     3961 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     4021 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     4081 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     4141 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     4201 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     4261 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     4321 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     4381 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     4441 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     4501 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     4561 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     4621 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     4681 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     4741 cttcagccat acttttcata ctcccgccat tcagag
//
