LOCUS       sequence-344648-        6387 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 6387)
  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 6387)
  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-NPC1_STOP
FEATURES             Location/Qualifiers
     source          1..6387
                     /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(3944..4043)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4060..4079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4061..4079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4084..4100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4084..4100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4202..4221)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4213..5022
                     /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(4280..4299)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5169..5757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5658..5677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5911..5928
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6087..6114)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6206..6292)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6341..6363
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6355..6372
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6356..6372
                     /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 ccaccatgac cgcaagggga
      121 ctggccctgg gactgctgct gctgctgctg tgcccagccc aggtgttctc tcagagctgc
      181 gtgtggtatg gcgagtgtgg catcgcctac ggcgacaagc ggtacaactg cgagtacagc
      241 ggaccaccta agccactgcc aaaggacggc tacgatctgg tgcaggagct gtgccctggc
      301 ttctttttcg gcaacgtgtc cctgtgctgt gacgtgcggc agctgcagac cctgaaggat
      361 aatctgcagc tgcctctgca gtttctgtct agatgcccaa gctgttttta taacctgctg
      421 aatctgttct gcgagctgac ctgttcccca aggcagtctc agttcctgaa cgtgaccgcc
      481 acagaggact acgtggatcc cgtgacaaac cagaccaaga caaatgtgaa ggagctgcag
      541 tactatgtgg gccagagctt cgccaacgcc atgtataatg catgcaggga cgtggaggca
      601 ccaagctcca atgataaggc cctgggactg ctgtgcggca aggacgcaga tgcctgtaac
      661 gccaccaatt ggatcgagta catgtttaac aaggataatg gccaggcccc tttcaccatc
      721 acaccagtgt tttccgactt ccccgtgcac ggcatggagc ctatgaacaa tgccaccaag
      781 ggctgcgacg agtctgtgga tgaggtgaca gccccttgca gctgtcagga ttgctccatc
      841 gtgtgcggac ctaagccaca gccaccacct ccaccagcac catggaccat cctgggactg
      901 gacgccatgt atgtgatcat gtggatcaca tacatggcct ttctgctggt gtttttcggc
      961 gcctttttcg ccgtgtggtg ctatcggaag agatacttcg tgagcgagta cacccccatc
     1021 gacagcaaca tcgccttcag cgtgaatgcc tccgacaagg gcgaggcatc ctgctgtgat
     1081 cccgtgagcg ccgccttcga gggatgcctg cggagactgt ttacccggtg gggctctttc
     1141 tgcgtgagaa acccaggctg cgtgatcttt ttcagcctgg tgtttatcac agcctgttct
     1201 agcggcctgg tgttcgtgag ggtgaccaca aatccagtgg atctgtggag cgccccatcc
     1261 tctcaggcaa ggctggagaa ggagtacttt gaccagcact tcggcccatt tttcaggacc
     1321 gagcagctga tcatcagggc accactgaca gataagcaca tctaccagcc atatcctagc
     1381 ggagcagacg tgcccttcgg ccctccactg gatatccaga tcctgcacca ggtgctggac
     1441 ctgcagatcg ccatcgagaa catcaccgcc tcctatgaca atgagaccgt gacactgcag
     1501 gatatctgcc tggcccctct gagcccatat aacaccaatt gtacaatcct gtccgtgctg
     1561 aactactttc agaattccca ctctgtgctg gatcacaaga agggcgacga cttcttcgtg
     1621 tacgccgact atcacaccca cttcctgtac tgcgtgcggg cccctgcctc tctgaacgac
     1681 accagcctgc tgcacgatcc atgtctgggc acatttggcg gacccgtgtt cccttggctg
     1741 gtgctgggcg gctatgacga tcagaactac aacaatgcca ccgccctggt catcaccttc
     1801 cctgtgaaca attactataa tgataccgag aagctgcaga gagcccaggc ctgggagaag
     1861 gagtttatca acttcgtgaa gaattacaag aacccaaatc tgaccatcag cttcacagcc
     1921 gagcggtcca tcgaggacga gctgaacaga gagagcgact ccgacgtgtt caccgtggtc
     1981 atcagctatg ccatcatgtt cctgtacatc agcctggccc tgggccacat gaagtcctgc
     2041 aggcgcctgc tggtggatag caaggtgtcc ctgggcatcg ccggcatcct gatcgtgctg
     2101 agctccgtgg cctgttccct gggcgtgttc tcttatatcg gcctgcctct gaccctgatc
     2161 gtgatcgaag tgatcccatt tctggtgctg gccgtgggcg tggacaatat cttcatcctg
     2221 gtgcaggcat accagaggga cgagcgcctg cagggagaga cactggatca gcagctggga
     2281 agggtgctgg gagaggtggc accatccatg tttctgtcta gcttctctga gaccgtggcc
     2341 tttttcctgg gcgccctgtc cgtgatgcca gcagtgcaca cattttctct gttcgccggc
     2401 ctggccgtgt ttatcgactt cctgctgcag atcacctgct tcgtgagcct gctgggcctg
     2461 gacatcaaga ggcaggagaa gaaccgcctg gatatcttct gctgcgtgcg gggagcagag
     2521 gacggcacat ccgtgcaggc ctctgagagc tgtctgttcc ggtttttcaa gaattcctat
     2581 tctccactgc tgctgaagga ctggatgaga cccatcgtga tcgccatctt tgtgggcgtg
     2641 ctgagcttct ccatcgccgt gctgaacaag gtggacatcg gcctggatca gtctctgagc
     2701 atgcccgacg atagctatat ggtggattac tttaagtcca tctctcagta tctgcacgca
     2761 ggaccacccg tgtacttcgt gctggaggag ggccacgact acacctcctc taagggccag
     2821 aatatggtgt gcggcggcat gggctgtaac aatgattccc tggtgcagca gatcttcaac
     2881 gccgcccagc tggacaatta tacacggatc ggctttgccc ccagctcctg gatcgacgat
     2941 tacttcgatt gggtgaagcc tcagtctagc tgctgtagag tggacaacat caccgatcag
     3001 ttttgcaatg caagcgtggt ggaccctgca tgcgtgaggt gtaggccact gacaccagag
     3061 ggcaagcaga ggccacaggg aggcgacttc atgcgcttcc tgcctatgtt cctgtccgac
     3121 aacccaaatc caaagtgtgg caagggagga cacgcagcat actcctctgc cgtgaacatc
     3181 ctgctgggac acggaaccag ggtgggagcc acatacttca tgacctacca cacagtgctg
     3241 cagacctccg ccgacttcat cgatgccctg aagaaggcca ggctgatcgc ctctaacgtg
     3301 accgagacaa tgggcatcaa tggctctgcc tatcgcgtgt ttccctacag cgtgttttac
     3361 gtgttctatg agcagtacct gaccatcatc gacgatacaa tctttaacct gggcgtgagc
     3421 ctgggcgcca tcttcctggt gaccatggtg ctgctgggat gcgagctgtg gtccgccgtg
     3481 atcatgtgcg ccacaatcgc catggtgctg gtgaatatgt tcggcgtgat gtggctgtgg
     3541 ggcatcagcc tgaacgccgt gtccctggtg aacctggtca tgtcttgcgg catcagcgtg
     3601 gagttttgta gccacatcac cagagccttc acagtgtcca tgaagggatc tagggtggag
     3661 agagcagagg aggccctggc acacatggga agctccgtgt tttctggcat caccctgaca
     3721 aagttcggcg gcatcgtggt gctggccttt gccaagagcc agatcttcca gatcttttat
     3781 ttccggatgt acctggcaat ggtgctgctg ggagcaaccc acggcctgat ctttctgccc
     3841 gtgctgctgt cttatatcgg ccctagcgtg aacaaggcca agtcctgcgc caccgaggag
     3901 agatacaagg gcacagagcg ggagagactg ctgaatttct gagacccagc tttcttgtac
     3961 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     4021 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     4081 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     4141 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     4201 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     4261 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     4321 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     4381 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     4441 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     4501 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     4561 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     4621 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     4681 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     4741 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     4801 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     4861 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     4921 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     4981 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     5041 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     5101 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5161 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5221 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5281 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5341 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5401 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5461 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5521 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5581 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5641 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5701 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5761 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5821 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5881 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5941 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6001 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     6061 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     6121 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     6181 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     6241 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     6301 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     6361 acgacggcca gtcttaagct cgggccc
//
