LOCUS       sequence-479868-        4854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose-inducible plasmid with thermostable GFP (TGP) and 
            Twin-Strep tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4854)
  AUTHORS   Reddy KD, Rasool B, Akher FB, Kutlesic N, Pant S, Boudker O
  TITLE     Evolutionary analysis reveals the origin of sodium coupling in 
            glutamate transporters.
  JOURNAL   Nat Struct Mol Biol (2025).
  PUBMED    38106174
REFERENCE   2  (bases 1 to 4854)
  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: "Nat Struct 
            Mol Biol (2025)."
COMMENT     Sequence Label: pBAD24-TGP-TwinStrep
FEATURES             Location/Qualifiers
     source          1..4854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    149..289
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(215..234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     375..397
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(553..1431)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1458..1742
                     /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     1665..1684
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1801..1818
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(2515..2520,2521..2544,2545..2580,2581..2604)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             2629..2658
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2712..2729)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2882..2968
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3060..3087
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3106..3197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3198..3266,3267..4058)
                     /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(3416..3435)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4229..4817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4718..4737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
       61 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      121 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
      181 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      241 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
      301 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      361 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      421 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca
      481 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
      541 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
      601 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
      661 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
      721 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
      781 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
      841 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
      901 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
      961 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     1021 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     1081 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     1141 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     1201 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     1261 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     1321 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     1381 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     1441 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     1501 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     1561 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     1621 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     1681 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     1741 atacccgttt ttttgggcta gcaggaggaa ttcaccatgg tacccccggg ctcgagctcc
     1801 ctggtgccgc gtggtagcgg tggcggtagc ggcggtagcg cgagcgttat caagccggag
     1861 atgaagatta aactgcgtat ggaaggcgcg gtgaacggtc acaagttcgt tatcgagggt
     1921 gaaggcattg gtaaaccgta cgagggcacc cagaccctgg atctgaccgt ggaggaaggt
     1981 gcgccgctgc cgtttagcta cgacatcctg accccggcgt tccaatatgg taaccgtgcg
     2041 tttaccaagt acccggagga cattccggat tatttcaaac aggcgtttcc ggagggttac
     2101 agctgggaac gtagcatgac ctatgaagat caaggcatct gcattgcgac cagcgatatc
     2161 acgatggagg gtgactgctt cttttacgaa attcgtttcg acggcaccaa ctttccgccg
     2221 aacggtccgg tgatgcagaa gaaaaccctg aagtgggagc cgagcaccga aaaaatgtat
     2281 gtggaagatg gcgttctgaa aggtgacgtt gagatggcgc tgctgctgga aggcggtggc
     2341 cactaccgtt gcgatttcaa gaccacctat aaagcgaaga aagacgtgcg tctgccggat
     2401 gcgcacgagg ttgaccaccg tatcgaaatt ctgagccacg acaaggatta caacaaagtt
     2461 cgtctgtatg agcacgcgga agcgcgttac agcggtggcg gtagcggcgg tggcagcgcg
     2521 tggagccacc cgcagtttga gaagggtggc ggtagcggcg gtggcagcgg tggcagcgct
     2581 tggtctcacc cgcaatttga aaaaggtggc ggtagcggcg gtggcagcca tcatcaccat
     2641 caccaccatc accatcattg ataatctaga tctaagcttg gctgttttgg cggatgagag
     2701 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     2761 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     2821 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     2881 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgcc
     2941 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     3001 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     3061 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     3121 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3181 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3241 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3301 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3361 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3421 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     3481 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3541 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3601 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3661 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3721 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3781 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3841 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3901 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3961 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4021 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4081 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4141 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gcct
//
