LOCUS       sequence-163523-        5429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 485, Emission:
            507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5429)
  AUTHORS   Pedelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS
  TITLE     Engineering and characterization of a superfolder green fluorescent 
            protein.
  JOURNAL   Nat Biotechnol. 2006 Jan;24(1):79-88. doi: 10.1038/nbt1172
  PUBMED    16369541
REFERENCE   2  (bases 1 to 5429)
  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: 
            "Nat Biotechnol"; date: "2006-01"; volume: "24"; issue: "1"; pages: 
            "79-88"
COMMENT     Sequence Label: sfGFP-pBAD
FEATURES             Location/Qualifiers
     source          1..5429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     179..201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     339..358
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(538..1416)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1443..1727
                     /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     1650..1669
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1754..1776
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1796..1813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1817..1849
                     /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             1862..1882
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1889..1891,1892..1894,1895..2608)
                     /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"
     primer_bind     complement(1934..1955)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2542..2563
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2654..2671)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2824..2910
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3002..3029
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3048..3139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3140..3208,3209..4000)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3358..3377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4042..4497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4129..4148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4339..4360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4608..5196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5097..5116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5350..5367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5382..5429,1..93)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 catctggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
       61 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      121 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      181 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagaaggag
      241 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
      301 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
      361 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
      421 tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg
      481 gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta
      541 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
      601 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
      661 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
      721 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
      781 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
      841 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
      901 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
      961 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     1021 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     1081 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     1141 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     1201 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     1261 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     1321 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     1381 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag
     1441 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
     1501 cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
     1561 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
     1621 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
     1681 ggatactacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgtttttttg
     1741 ggctagaaat aattttgttt aactttaaga aggagatata catatgcggg gttctcatca
     1801 tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta
     1861 cgagaacctg tacttccagg gctcgagcat ggtgagcaag ggcgaggagc tgttcaccgg
     1921 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg
     1981 cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac
     2041 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2101 cttcagccgc taccccgacc acatgaagcg ccacgacttc ttcaagtccg ccatgcccga
     2161 aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc
     2221 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2281 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacgt
     2341 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     2401 cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2461 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgtgc tgagcaaaga
     2521 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2581 tcacggcatg gacgagctgt acaagtaaga attcgaagct tggctgtttt ggcggatgag
     2641 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     2701 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     2761 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     2821 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     2881 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     2941 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     3001 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     3061 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3301 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3361 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     3421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3661 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4021 atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4081 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4141 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4201 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4261 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4321 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4381 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4441 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     4501 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4561 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4621 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4681 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4741 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4801 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4861 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4921 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4981 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5041 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5101 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5161 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5221 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5281 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5341 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     5401 cttacgcatc tgtgcggtat ttcacaccg
//
