LOCUS       sequence-259094-        4401 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 4401)
  AUTHORS   Sijben HJ, van Oostveen WM, Hartog PBR, Stucchi L, Rossignoli A, 
            Maresca G, Scarabottolo L, IJzerman AP, Heitman LH
  TITLE     Label-free high-throughput screening assay for the identification of
            norepinephrine transporter (NET/SLC6A2) inhibitors.
  JOURNAL   Sci Rep. 2021 Jun 10;11(1):12290. doi: 10.1038/s41598-021-91700-7.
  PUBMED    34112854
REFERENCE   2  (bases 1 to 4401)
  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: "Sci Rep."; 
            date: "2021-06-10"; volume: "11(1):12290. doi"; pages: " 
            10.1038/s41598-021-91700-7"
COMMENT     Sequence Label: pDONR221_SLC6A2
FEATURES             Location/Qualifiers
     source          1..4401
                     /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(2526..2625)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2642..2661)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2643..2661)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2666..2682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2666..2682)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2784..2803)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2795..3604
                     /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(2862..2881)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3751..4339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4240..4259
                     /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 accatgctgc tggccaggat gaaccctcag gtgcagccag agaacaatgg
      721 agcagacacc ggaccagagc agccactgcg ggccagaaag acagccgagc tgctggtggt
      781 gaaggagagg aatggagtgc agtgcctgct ggcaccaagg gacggcgatg cacagccaag
      841 agagacctgg ggcaagaaga tcgactttct gctgagcgtg gtgggcttcg ccgtggatct
      901 ggccaacgtg tggcggtttc cctacctgtg ctataagaat ggcggcggcg cctttctgat
      961 cccttacacc ctgttcctga tcatcgccgg catgccactg ttctacatgg agctggccct
     1021 gggccagtat aacagagagg gcgccgccac agtgtggaag atctgcccat tctttaaggg
     1081 cgtgggctac gccgtgatcc tgatcgccct gtatgtgggc ttctactata acgtgatcat
     1141 cgcctggtct ctgtactatc tgttcagctc ctttaccctg aatctgccct ggaccgactg
     1201 cggccacaca tggaacagcc ccaattgtac agatcctaag ctgctgaacg gcagcgtgct
     1261 gggcaatcac accaagtact ccaagtataa gttcacacct gccgccgagt tttatgagcg
     1321 gggcgtgctg cacctgcacg agtctagcgg catccacgat atcggcctgc cacagtggca
     1381 gctgctgctg tgcctgatgg tggtggtcat cgtgctgtac ttttccctgt ggaagggcgt
     1441 gaagacctct ggcaaggtgg tgtggatcac cgccacactg ccatatttcg tgctgtttgt
     1501 gctgctggtg cacggagtga cactgccagg agcctccaac ggcatcaatg cctacctgca
     1561 catcgacttc tataggctga aggaggccac cgtgtggatc gatgccgcca cacagatctt
     1621 ctttagcctg ggagcaggat tcggcgtgct gatcgccttt gcctcctaca acaagttcga
     1681 caacaattgc tatcgcgatg ccctgctgac ctcctctatc aattgtatca caagcttcgt
     1741 gtccggcttc gccatcttta gcatcctggg ctacatggcc cacgagcaca aagtgaatat
     1801 cgaggacgtg gcaaccgagg gagcaggact ggtgtttatc ctgtatcctg aggccatctc
     1861 caccctgtcc ggctctacat tctgggccgt ggtgttcttt gtgatgctgc tggccctggg
     1921 actggatagc tccatgggag gcatggaggc cgtgatcaca ggcctggccg acgattttca
     1981 ggtgctgaag aggcaccgca agctgttcac ctttggcgtg accttctcca catttctgct
     2041 ggccctgttc tgcatcacaa agggcggcat ctacgtgctg accctgctgg acacatttgc
     2101 cgccggcacc tctatcctgt tcgccgtgct gatggaggcc atcggcgtgt cttggtttta
     2161 cggcgtggac cggttcagca acgatatcca gcagatgatg ggctttaggc ccggcctgta
     2221 ttggcgcctg tgctggaagt tcgtgtcccc tgccttcctg ctgtttgtgg tggtggtgtc
     2281 tatcatcaac tttaagccac tgacctacga cgattatatc ttcccacctt gggcaaattg
     2341 ggtgggatgg ggaatcgccc tgtctagcat ggtgctggtg cccatctacg tgatctataa
     2401 gtttctgtct acccagggaa gcctgtggga gaggctggca tacggcatca cacctgagaa
     2461 tgagcaccac ctggtggccc agcgggatat cagacagttc cagctgcagc actggctggc
     2521 catcgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2581 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2641 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2701 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2761 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2821 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2881 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2941 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     3001 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3061 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3121 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3181 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3241 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3301 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3361 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3421 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3481 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3541 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3601 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3661 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3721 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3781 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3841 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3901 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3961 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4021 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4081 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4141 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4201 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4261 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4321 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4381 ctggcctttt gctcacatgt t
//
