LOCUS       sequence-257481-        4701 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 4701)
  AUTHORS   Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV
  TITLE     An enhanced monomeric blue fluorescent protein with the high 
            chemical stability of the chromophore.
  JOURNAL   PLoS One. 2011;6(12):e28674. Epub 2011 Dec 8.
  PUBMED    22174863
REFERENCE   2  (bases 1 to 4701)
  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: "PLoS One.";
            date: "2011"; volume: "6(12)"; pages: "e28674. Epub 2011 Dec 8"
COMMENT     Sequence Label: pBAD-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..4701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             43..60
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             70..771
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(817..834)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      987..1073
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1165..1192
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1211..1302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1303..1371,1372..2163)
                     /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(1521..1540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2334..2922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2823..2842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3076..3093
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3108..3248
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3174..3193)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3334..3356
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3512..4390)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4417..4701
                     /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     4624..4643
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 acccgttttt tgggctaaca ggaggaatta accatgggga gccaccatca ccatcaccat
       61 ggcagatcta tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
      121 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
      181 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
      241 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc
      301 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
      361 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
      421 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca
      481 ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga
      541 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacgc caagaccaca
      601 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac
      661 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca
      721 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatta agaattcgaa
      781 gcttggctgt tttggtggat gagagaagat tttcagcctg atacagatta aatcagaacg
      841 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
      901 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
      961 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1021 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     1081 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     1141 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     1201 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1261 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1321 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1381 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1441 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1501 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     1561 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1621 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1681 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1741 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1801 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1861 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1921 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1981 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2041 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2101 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2161 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2221 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2341 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2401 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2461 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2521 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2581 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2641 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2701 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2761 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2821 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2881 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2941 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3001 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3061 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3121 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     3181 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3241 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3301 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3361 caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc
     3421 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg
     3481 tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca
     3541 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     3601 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     3661 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     3721 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     3781 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     3841 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     3901 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     3961 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     4021 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     4081 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     4141 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     4201 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     4261 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     4321 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     4381 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     4441 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     4501 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     4561 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     4621 gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc
     4681 aactctctac tgtttctcca t
//
