LOCUS       pBad-sfGFP                4737 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Arabinose inducible E. coli codon optimized superfolder GFP with 
            C-terminal His6 tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-sfGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4737)
  AUTHORS   Miyake-Stoner SJ, Refakis CA, Hammill JT, Lusic H, Hazen JL, Deiters
            A, Mehl RA
  TITLE     Generating permissive site-specific unnatural aminoacyl-tRNA 
            synthetases.
  JOURNAL   Biochemistry. 2010 Mar 2;49(8):1667-77. doi: 10.1021/bi901947r.
  PUBMED    20082521
REFERENCE   2  (bases 1 to 4737)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..4737
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             319..1035
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             1042..1059
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1138..1155)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1308..1394
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1486..1513
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1532..1623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1624..2484
                     /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(1842..1861)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2655..3243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3144..3163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3397..3414
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3429..3569
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3495..3514)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3655..3677
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3833..4711)
                     /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 ggttagcaaa ggtgaagaac tgtttaccgg cgttgtgccg
      361 attctggtgg aactggatgg tgatgtgaat ggccataaat ttagcgttcg tggcgaaggc
      421 gaaggtgatg cgaccaacgg taaactgacc ctgaaattta tttgcaccac cggtaaactg
      481 ccggttccgt ggccgaccct ggtgaccacc ctgacctatg gcgttcagtg ctttagccgc
      541 tatccggatc atatgaaacg ccatgatttc tttaaaagcg cgatgccgga aggctatgtg
      601 caggaacgta ccattagctt caaagatgat ggcacctata aaacccgtgc ggaagttaaa
      661 tttgaaggcg ataccctggt gaaccgcatt gaactgaaag gtattgattt taaagaagat
      721 ggcaacattc tgggtcataa actggaatat aatttcaaca gccataatgt gtatattacc
      781 gccgataaac agaaaaatgg catcaaagcg aactttaaaa tccgtcacaa cgtggaagat
      841 ggtagcgtgc agctggcgga tcattatcag cagaataccc cgattggtga tggcccggtg
      901 ctgctgccgg ataatcatta tctgagcacc cagagcgttc tgagcaaaga tccgaatgaa
      961 aaacgtgatc atatggtgct gctggaattt gttaccgccg cgggcattac ccacggtatg
     1021 gatgaactgt ataaaggcag ccaccatcat catcaccatt aaagctcgag atctgcagct
     1081 ggtaccatat gggaattcga agcttggctg ttttggcgga tgagagaaga ttttcagcct
     1141 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     1201 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     1261 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     1321 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     1381 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     1441 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     1501 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     1561 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1621 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1681 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1741 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1801 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1861 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1921 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1981 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2041 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2101 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2161 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2221 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2281 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2341 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2401 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2461 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2521 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2641 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2701 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2761 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2821 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2881 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2941 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3001 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3061 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3301 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3361 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3421 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3481 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3541 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3601 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3661 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt
     3721 cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca aatggacgaa gcagggattc
     3781 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
     3841 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
     3901 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
     3961 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
     4021 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
     4081 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
     4141 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
     4201 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
     4261 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
     4321 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
     4381 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
     4441 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
     4501 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
     4561 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
     4621 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
     4681 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagag
//