LOCUS       sequence-261092-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5283)
  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 5283)
  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_SLC4A1
FEATURES             Location/Qualifiers
     source          1..5283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3408..3507)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3524..3543)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3525..3543)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3548..3564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3548..3564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3666..3685)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3677..4486
                     /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(3744..3763)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4633..5221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5122..5141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggagg agctgcagga cgattacgag gacatgatgg aggagaacct
      721 ggagcaggag gagtatgagg accccgatat ccctgagtcc cagatggagg agccagcagc
      781 acacgatacc gaggcaaccg ccacagacta ccacaccaca tctcaccccg gcacacacaa
      841 ggtgtatgtg gagctgcagg agctggtcat ggatgagaag aatcaggagc tgcggtggat
      901 ggaggccgcc agatgggtgc agctggagga gaacctggga gagaatggag catggggcag
      961 accccacctg tcccacctga cattttggtc tctgctggag ctgcggagag tgttcaccaa
     1021 gggcacagtg ctgctggacc tgcaggagac ctctctggca ggagtggcaa accagctgct
     1081 ggacaggttt atcttcgagg atcagatcag gcctcaggac agggaggagc tgctgcgcgc
     1141 cctgctgctg aagcactccc acgcaggaga gctggaggcc ctgggaggcg tgaagccagc
     1201 cgtgctgaca agatctggcg atccaagcca gccactgctg cctcagcaca gctccctgga
     1261 gacccagctg ttctgcgagc agggcgacgg aggaacagag ggacactccc catctggcat
     1321 cctggagaag atcccccctg atagcgaggc caccctggtg ctggtgggaa gggcagactt
     1381 tctggagcag cccgtgctgg gcttcgtgag actgcaggag gcagcagagc tggaggcagt
     1441 ggagctgcca gtgcctatcc ggtttctgtt cgtgctgctg ggaccagagg caccacacat
     1501 cgattacacc cagctgggaa gagcagcagc cacactgatg agcgagcggg tgttcagaat
     1561 cgacgcctat atggcacagt ccaggggaga gctgctgcac tctctggagg gctttctgga
     1621 ttgttccctg gtgctgccac caaccgacgc accaagcgag caggccctgc tgtccctggt
     1681 gccagtgcag agggagctgc tgaggcgccg gtaccagtct agccctgcca agccagattc
     1741 ctctttctat aagggactgg acctgaatgg aggaccagac gatcctctgc agcagacagg
     1801 ccagctgttt ggcggcctgg tgagggacat cagaaggcgc tacccttact atctgagcga
     1861 catcaccgat gccttctccc cacaggtgct ggcagccgtg atctttatct atttcgccgc
     1921 cctgagccca gcaatcacct ttggcggact gctgggcgag aagacacgca accagatggg
     1981 cgtgtctgag ctgctgatca gcacagccgt gcagggcatc ctgttcgcac tgctgggagc
     2041 acagccactg ctggtggtgg gcttcagcgg ccctctgctg gtgtttgagg aggccttctt
     2101 ttccttctgc gagaccaatg gcctggagta catcgtgggc agggtgtgga tcggcttctg
     2161 gctgatcctg ctggtggtgc tggtggtggc ctttgagggc tctttcctgg tgcggtttat
     2221 cagcagatac acccaggaga tcttttcctt cctgatctct ctgatcttca tctatgagac
     2281 cttcagcaag ctgatcaaga tcttccagga tcaccccctg cagaagacct acaactataa
     2341 tgtgctgatg gtgccaaagc cacagggacc tctgccaaac accgccctgc tgagcctggt
     2401 gctgatggca ggcacattct ttttcgccat gatgctgcgc aagttcaaga atagctccta
     2461 ctttcccggc aagctgcgga gagtgatcgg cgatttcggc gtgcctatca gcatcctgat
     2521 catggtgctg gtggactttt tcatccagga tacctataca cagaagctgt ccgtgcctga
     2581 cggctttaag gtgtctaact ctagcgccag gggatgggtc atccaccctc tgggactgcg
     2641 ctccgagttc ccaatctgga tgatgtttgc ctctgccctg cctgccctgc tggtgttcat
     2701 cctgatcttt ctggagagcc agatcaccac actgatcgtg tccaagccag agcggaagat
     2761 ggtgaagggc tccggctttc acctggacct gctgctggtg gtgggaatgg gaggagtggc
     2821 cgccctgttc ggaatgccat ggctgtctgc caccacagtg agaagcgtga cccacgccaa
     2881 tgccctgaca gtgatgggca aggcatctac ccctggagca gcagcacaga tccaggaggt
     2941 gaaggagcag aggatcagcg gactgctggt ggccgtgctg gtgggactgt ctatcctgat
     3001 ggagcctatc ctgagccgca tcccactggc cgtgctgttt ggcatcttcc tgtacatggg
     3061 cgtgaccagc ctgtccggca tccagctgtt tgataggatc ctgctgctgt tcaagcctcc
     3121 aaagtaccac ccagacgtgc cctatgtgaa gagggtgaag acctggcgca tgcacctgtt
     3181 cacaggcatc cagatcatct gcctggccgt gctgtgggtg gtgaagagca ccccagcatc
     3241 cctggccctg ccttttgtgc tgatcctgac agtgccactg aggcgcgtgc tgctgccact
     3301 gatcttccgg aacgtggagc tgcagtgtct ggatgccgac gatgccaagg ccacctttga
     3361 cgaggaggag ggcagagacg agtacgatga ggtggccatg cccgtggacc cagctttctt
     3421 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     3481 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     3541 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     3601 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     3661 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     3721 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     3781 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3841 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3901 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3961 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     4021 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     4081 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     4141 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     4201 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     4261 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     4321 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     4381 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     4441 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     4501 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     4561 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     4621 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4681 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4741 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4801 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4861 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4921 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4981 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5041 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5101 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5161 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5221 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5281 gtt
//
