LOCUS       sequence-390324-        4350 bp    DNA     circular SYN 14-APR-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 4350)
  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 4350)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-SLC6A1_STOP
FEATURES             Location/Qualifiers
     source          1..4350
                     /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(1907..2006)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2023..2042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2024..2042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2047..2063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2047..2063)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2165..2184)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2176..2985
                     /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(2243..2262)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3132..3720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3621..3640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3874..3891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4050..4077)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4169..4255)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4304..4326
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4319..4335
                     /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 ccaccatggc cacaaacggc
      121 tccaaggtgg ccgacggcca gatctccacc gaggtgtctg aggcccctgt ggccaatgat
      181 aagccaaaga cactggtggt gaaggtgcag aagaaggcag cagacctgcc tgacagggat
      241 acctggaagg gcagattcga ttttctgatg tcttgcgtgg gatacgcaat cggactggga
      301 aacgtgtggc ggttcccata cctgtgcggc aagaatggcg gcggcgcctt tctgatccct
      361 tacttcctga cactgatctt tgccggcgtg ccactgtttc tgctggagtg cagcctgggc
      421 cagtatacct ccatcggagg actgggcgtg tggaagctgg cacccatgtt caagggagtg
      481 ggactggcag cagccgtgct gtccttttgg ctgaacatct actatatcgt gatcatctct
      541 tgggccatct actatctgta caatagcttc accacaaccc tgccatggaa gcagtgcgac
      601 aacccctgga atacagatag gtgtttctcc aactattcta tggtgaacac aaccaatatg
      661 acaagcgccg tggtggagtt ttgggagagg aatatgcacc agatgaccga cggcctggat
      721 aagccaggac agatcaggtg gcctctggca atcaccctgg ccatcgcctg gatcctggtg
      781 tacttctgca tctggaaggg cgtgggctgg acaggcaagg tggtgtattt ttccgccacc
      841 tacccttata tcatgctgat catcctgttc tttaggggag tgacactgcc aggagcaaag
      901 gagggcatcc tgttctacat cacccctaac tttagaaagc tgtccgactc tgaagtgtgg
      961 ctggatgccg ccacccagat cttcttttcc tatggactgg gactgggatc tctgatcgcc
     1021 ctgggaagct acaactcctt ccacaacaac gtgtacaggg actctatcat cgtgtgctgt
     1081 atcaattctt gtacaagcat gttcgccggc tttgtgatct tcagcatcgt gggcttcatg
     1141 gcccacgtga ccaagcgcag catcgcagat gtggcagcat ccggaccagg actggccttt
     1201 ctggcatacc ctgaggcagt gacccagctg cccatctccc ctctgtgggc catcctgttc
     1261 ttttctatgc tgctgatgct gggcatcgac agccagttct gcacagtgga gggctttatc
     1321 accgccctgg tggatgagta tcccaggctg ctgcgcaacc ggagagagct gttcatcgcc
     1381 gccgtgtgca tcatctctta cctgatcggc ctgagcaata tcacacaggg cggcatctac
     1441 gtgttcaagc tgttcgacta ctatagcgcc tccggcatgt ccctgctgtt cctggtgttc
     1501 tttgagtgcg tgagcatcag ctggttctac ggcgtgaaca ggttttatga taatatccag
     1561 gagatggtgg gctcccgccc atgcatctgg tggaagctgt gctggtcttt ctttaccccc
     1621 atcatcgtgg ccggcgtgtt catctttagc gccgtgcaga tgacaccact gaccatgggc
     1681 aactacgtgt tcccaaagtg gggacaggga gtgggatggc tgatggcact gagctccatg
     1741 gtgctgatcc ccggctacat ggcctatatg tttctgacac tgaagggcag cctgaagcag
     1801 cggatccagg tcatggtgca gccttccgag gacatcgtga gaccagagaa tggaccagag
     1861 cagccacagg caggatctag caccagcaag gaggcctaca tctaagaccc agctttcttg
     1921 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1981 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     2041 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2101 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2161 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2221 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2281 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2341 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2401 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2461 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2521 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     2581 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     2641 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2701 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2761 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     2821 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     2881 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     2941 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     3001 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     3061 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3121 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3181 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3241 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3301 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3361 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3421 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3481 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3541 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3601 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3661 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3721 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3781 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3841 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3901 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3961 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     4021 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     4081 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     4141 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     4201 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     4261 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     4321 aaaacgacgg ccagtcttaa gctcgggccc
//
