LOCUS       sequence-353417-        4440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4440)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 4440)
  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 Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221-SLC7A1_STOP
FEATURES             Location/Qualifiers
     source          1..4440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1997..2096)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2113..2132)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2114..2132)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2137..2153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2137..2153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2255..2274)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2266..3075
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2333..2352)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3222..3810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3711..3730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3964..3981
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4140..4167)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4259..4345)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4394..4416
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4408..4425
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4409..4425
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggg ctgcaaggtg
      121 ctgctgaaca tcggccagca gatgctgcgg agaaaggtgg tggactgcag cagggaggag
      181 accaggctgt ccaggtgtct gaatacattt gatctggtgg ccctgggagt gggaagcacc
      241 ctgggagccg gcgtgtatgt gctggcagga gcagtggcaa gggagaacgc aggccctgcc
      301 atcgtgatct cttttctgat cgccgccctg gcaagcgtgc tggcaggact gtgctacgga
      361 gagttcggag caagggtgcc aaagacagga tccgcctacc tgtattctta cgtgaccgtg
      421 ggcgagctgt gggccttcat cacaggctgg aatctgatcc tgtcctatat catcggcacc
      481 agctccgtgg caagggcatg gtctgccaca tttgacgagc tgatcggccg gcctatcggc
      541 gagttctcta gaacccacat gacactgaac gcccctggcg tgctggccga gaatccagat
      601 atctttgccg tgatcatcat cctgatcctg accggcctgc tgacactggg cgtgaaggag
      661 tccgccatgg tgaacaagat cttcacctgc atcaatgtgc tggtgctggg ctttatcatg
      721 gtgtctggct tcgtgaaggg cagcgtgaag aactggcagc tgaccgagga ggactttggc
      781 aatacaagcg gccggctgtg cctgaacaat gataccaagg agggcaagcc aggagtggga
      841 ggcttcatgc ctttcggctt ttccggcgtg ctgtctggag cagcaacatg cttctacgcc
      901 tttgtgggct tcgactgtat cgccaccaca ggcgaggagg tgaagaaccc ccagaaggcc
      961 atccctgtgg gcatcgtggc ctctctgctg atctgcttta tcgcctattt cggcgtgagc
     1021 gccgccctga ccctgatgat gccatacttt tgtctggaca acaatagccc cctgcctgat
     1081 gccttcaagc acgtgggatg ggagggagca aagtatgcag tggcagtggg aagcctgtgc
     1141 gcactgtctg ccagcctgct gggatccatg ttcccaatgc ccagagtgat ctacgccatg
     1201 gccgaggacg gcctgctgtt caagtttctg gccaacgtga atgatcgcac caagacaccc
     1261 atcatcgcca ccctggcatc cggagcagtg gcagccgtga tggccttcct gtttgacctg
     1321 aaggatctgg tggacctgat gtccatcggc acactgctgg cctattctct ggtggcagca
     1381 tgcgtgctgg tgctgaggta tcagcctgag cagccaaatc tggtgtacca gatggcctcc
     1441 acctctgatg agctggaccc agccgatcag aacgagctgg ccagcacaaa tgactcccag
     1501 ctgggctttc tgcccgaggc cgagatgttc tccctgaaga ccatcctgtc tccaaagaac
     1561 atggagccct ctaagatcag cggcctgatc gtgaatatca gcacatccct gatcgccgtg
     1621 ctgatcatca ccttttgcat cgtgacagtg ctgggaaggg aggccctgac caagggcgcc
     1681 ctgtgggccg tgttcctgct ggcaggaagc gccctgctgt gcgccgtggt gaccggcgtg
     1741 atctggagac agcccgagag caagacaaag ctgtccttta aggtgccctt cctgcctgtg
     1801 ctgccaatcc tgtccatctt tgtgaacgtg tacctgatga tgcagctgga tcagggaacc
     1861 tgggtgcggt tcgccgtgtg gatgctgatt ggctttatca tctatttcgg ctacggcctg
     1921 tggcactctg aggaggcaag cctggacgca gatcaggcaa gaacacctga cggcaatctg
     1981 gatcagtgca agtaagaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     2041 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     2101 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     2161 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     2221 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     2281 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2341 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     2401 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2461 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2521 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     2581 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2641 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2701 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2761 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2821 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2881 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2941 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     3001 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     3061 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     3121 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     3181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3841 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3901 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3961 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4021 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4081 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     4141 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     4201 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     4261 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     4321 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     4381 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
