LOCUS       Exported                4733 bp ds-DNA     circular SYN 26-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBAD_FLAG-GFP(Y38TAG)-6His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4733)
  AUTHORS   Christine
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 1, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Freezer Position: -20*C Plasmid Box #5: 13-15; -80* Plasmid Box #2 :
            67
FEATURES             Location/Qualifiers
     source          1..4733
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    252..1013
                     /label=Flag-GFP (Y39TAG)-6His
                     /label=Flag-GFP\(Y39TAG)-6His
                     /note="/vntifkey=21"
     CDS             255..278
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             279..992
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT*GKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             993..1010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1236..1322
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1414..1441
                     /gene="rrnB_T2 term"
                     /label=rrnB_T2\term
                     /note="Terminator"
                     /note="/vntifkey=43"
     promoter        1461..1552
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1553..2413
                     /codon_start=1
                     /label=ampR
                     /note="ampR"
                     /note="/sequence=ORF_1 rf(3)"
                     /note="/vntifkey=4"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2584..3172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3358..3498
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3762..4640)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(4667..4733,1..218)
                     /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)"
ORIGIN
        1 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
       61 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
      121 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
      181 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttg ggctaacagg
      241 aggaattaac catggattac aaagatgatg atgataaagt gagcaagggc gaggagctgt
      301 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      361 gcgtgtccgg cgagggcgag ggcgatgcca cctagggcaa gctgaccctg aagttcatct
      421 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      481 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      541 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      601 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      661 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
      721 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
      781 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
      841 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
      901 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
      961 ggatcactct cggcatggac gagctgtaca agcatcacca tcaccatcac tgagcggccg
     1021 cactcgagag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1081 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1141 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1201 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1261 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1321 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1381 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1441 ttgcgtttct acaaactctt tttgtttatt tttctaaata cattcaaata tgtatccgct
     1501 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1561 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1621 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1681 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1741 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     1801 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1861 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1921 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1981 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2041 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2101 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2161 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2221 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2281 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2341 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2401 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2461 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2521 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2581 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2641 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2701 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2761 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2821 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2881 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2941 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3001 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3061 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3121 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3181 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3241 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3301 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3361 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     3421 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3481 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3541 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3601 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag
     3661 gcgaagcggc atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta
     3721 tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct
     3781 acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat
     3841 tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg
     3901 gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg
     3961 cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc
     4021 gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg
     4081 ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta
     4141 atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa
     4201 tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc
     4261 tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc
     4321 cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc
     4381 tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt
     4441 cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg
     4501 agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc
     4561 tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga
     4621 tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt
     4681 ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc gct
//
