LOCUS       sequence-440990-        6999 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 6999)
  TITLE     Human Transporter Knockout library
REFERENCE   2  (bases 1 to 6999)
  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"
COMMENT     Sequence Label: pENTR-ABCC1_K684I_STOP
FEATURES             Location/Qualifiers
     source          1..6999
                     /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"
     regulatory      101..110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4735..4834)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4891..4907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4891..4907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4904..4926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(5079..5667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5159..5178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5803..6618)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5803..6618)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6526..6545
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6610..6629
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     6739..6755
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6739..6755
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      6756..6802
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6916..6938
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6930..6947
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6931..6947
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacagat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct gccaccatgg cactccgagg
      121 cttttgtagc gcagatggca gcgaccccct gtgggattgg aatgtaacat ggaatacaag
      181 caaccctgac ttcacaaaat gttttcaaaa tacagtactc gtctgggtcc cctgcttcta
      241 cttgtgggca tgtttcccct tctacttctt gtatctctcc aggcacgata ggggatacat
      301 ccaaatgacc cctctgaata aaaccaaaac tgctctgggc ttcctgttat ggatagtctg
      361 ctgggccgac ttattttatt ccttttggga gcgctcaagg ggcatcttcc tcgcccccgt
      421 tttccttgta tctcctacgt tgcttggaat cactatgtta ttagctactt tcctgatcca
      481 actcgaaaga agaaaagggg ttcaaagcag tggaataatg ctcacctttt ggctcgtagc
      541 cttggtgtgc gcactcgcaa tcttgaggtc aaagatcatg actgcactga aggaagatgc
      601 ccaagtagat ttgtttcgcg acataacttt ctacgtgtat ttctcacttc ttctgatcca
      661 actggtcctt tcttgcttct ccgatcggag ccctttattt tccgagacca tccatgaccc
      721 aaacccttgt ccagagagct ctgcgagttt cttatctagg atcaccttct ggtggatcac
      781 cggtttaatc gttcgcggtt atcggcagcc gcttgagggc tctgaccttt ggagtcttaa
      841 caaagaggat acgtcagaac aagtggttcc tgtactggtg aagaattgga agaaagaatg
      901 tgccaaaact cgcaagcaac ccgttaaagt agtgtattct agcaaggatc ccgcccagcc
      961 gaaagagtca tccaaagtgg atgccaatga ggaagttgaa gcattgattg taaagtctcc
     1021 tcagaaagag tggaaccctt cattgtttaa ggttctgtat aagacatttg gtccctactt
     1081 tctgatgtct ttcttcttca aagcgatcca tgaccttatg atgttttcag gtccccaaat
     1141 tctgaagtta cttattaagt ttgtgaatga taccaaggca ccggattggc aggggtattt
     1201 ctacactgtt ctcctgtttg taacagcgtg ccttcaaaca ctcgtcttac accagtattt
     1261 ccacatatgc ttcgtgtcag gaatgaggat taagacagcc gttatcggtg ctgtctacag
     1321 aaaggctctc gtcatcacta acagcgcgag aaaatcttcc acagtgggtg aaattgtgaa
     1381 cctgatgagc gtggatgcgc aaagatttat ggacctcgcc acctatatta acatgatatg
     1441 gagcgcaccc ctgcaggtca tattggcact ctacttgctt tggttgaacc tgggtccaag
     1501 tgttctggct ggtgttgctg tgatggtgtt gatggtaccc gttaacgccg tcatggccat
     1561 gaagactaag acatatcaag tcgctcatat gaaatccaag gacaaccgaa taaaactcat
     1621 gaacgaaatc ctgaatggta ttaaagtgct taaactgtat gcttgggaac tggctttcaa
     1681 agataaggtc ttggctatcc gtcaagaaga attgaaagtt cttaagaaat cagcttacct
     1741 ctcagccgtc ggcacgttca cctgggtctg cacacccttt ctggtggcct tgtgtacttt
     1801 cgccgtatat gtcactattg acgagaacaa tatcctggac gcacaaaccg cctttgtatc
     1861 cctcgcgctg tttaacattc tccggtttcc gcttaatatc ttgcctatgg tgatctcaag
     1921 tattgtccaa gccagtgtct ctctgaaacg gctccgcatc tttctgtctc atgaggagtt
     1981 ggaaccagat tctatagaac gccgacccgt gaaagacggc ggcgggacca acagcataac
     2041 tgtcagaaac gcaacattca catgggctcg cagtgacccg cccaccctca atggcattac
     2101 attctccatt ccggaaggcg cattagtcgc agtggtgggg caagttggat gtggcatatc
     2161 cagcctgctc tccgctctgc ttgccgagat ggacaaagtt gagggacatg tcgctataaa
     2221 agggtctgtc gcatatgtcc cccaacaagc ctggatacag aatgattctt tacgagagaa
     2281 catattgttt ggatgtcaac tcgaagaacc ctattaccgg tccgttattc aagcatgtgc
     2341 tctcttgcct gacctcgaga ttcttcccag cggcgaccgt acagagatcg gcgagaaagg
     2401 cgtcaatctc tctggtggcc aaaagcagcg cgtcagcttg gcacgtgccg tgtattctaa
     2461 tgctgatatt tatctgttcg atgatccact ctctgccgtg gatgcccatg tggggaaaca
     2521 catattcgag aatgtaatag ggcccaaggg tatgcttaag aataaaacta ggattctggt
     2581 aacccatagt atgtcttatt tgccacaagt tgatgtcatt atcgtcatgt ctggcggaaa
     2641 gattagtgag atgggctctt accaagaact gctggcccgg gacggggcct ttgccgaatt
     2701 cctgcgtact tacgcctcaa cggaacagga gcaggatgct gaggagaatg gagtaaccgg
     2761 agttagcggg ccaggtaaag aggctaaaca aatggagaac gggatgttag tcactgactc
     2821 tgcaggaaaa caactgcaac gccagctgag cagcagcagt agctattccg gcgacatttc
     2881 ccgacaccac aactctactg cggaacttca gaaagcggaa gctaagaaag aggaaacttg
     2941 gaaactgatg gaagctgata aggctcaaac cgggcaagtg aaactgtctg tttattggga
     3001 ttatatgaag gccattgggt tgtttattag cttcctgtca attttcctgt tcatgtgtaa
     3061 tcatgtttca gctttagcct ctaactattg gctttctctc tggaccgatg atcctattgt
     3121 taatggtact caagagcata ctaaagtaag gctcagcgtg tatggtgccc tgggcataag
     3181 ccaaggtatc gccgtatttg ggtacagcat ggctgtatcc atcggtggaa tcctcgcgtc
     3241 acgctgcctt catgtcgact tgttacattc aatactgagg agtcctatgt ccttctttga
     3301 aagaacacct agcggaaacc tggtcaaccg attttcaaag gaactggata ctgtcgactc
     3361 catgattcct gaggttatta aaatgtttat gggctcttta tttaacgtga ttggagcctg
     3421 tatcgtcatc ctgcttgcca cgccaatcgc agcgataatc attccacccc tgggacttat
     3481 ctacttcttt gtgcaacgct tctacgtggc gtcatcccga cagctcaaac gcctggaaag
     3541 cgtctccaga agcccagtgt actcacattt caacgagacc ctgcttgggg tttccgtgat
     3601 aagggctttc gaggaacagg aacgctttat acatcaatcc gacttgaaag tggatgagaa
     3661 ccaaaaggcc tattatccgt ccatagtagc aaacaggtgg ctggctgtac gtctggagtg
     3721 cgtgggaaat tgcattgttc tgtttgcagc tttatttgcc gttatatcca ggcacagtct
     3781 ctcagccggg ctcgttgggt tgtctgtcag ctactcactg caagtgacga cttatcttaa
     3841 ttggcttgtg cgcatgagct cagaaatgga aacaaatatc gtagcagttg aacggcttaa
     3901 agaatactca gaaacagaga aagaggcacc ttggcaaata caagaaactg caccaccttc
     3961 cagttggcct caagtcggcc gggtcgagtt cagaaattat tgtctgaggt atcgagaaga
     4021 cctggacttt gtgctgagac atattaatgt taccattaat ggcggtgaaa aggtcgggat
     4081 tgtaggaagg actggtgcag gtaaatccag ccttacgtta ggccttttcc gtatcaacga
     4141 atccgctgaa ggcgagatca taatagatgg gattaatata gccaaaatcg gccttcatga
     4201 tcttcgcttc aagatcacta tcatccctca ggaccccgtt ttgttttctg gaagcttacg
     4261 catgaacttg gatccattct cacagtattc cgacgaagaa gtgtggactt cactcgaatt
     4321 ggcgcacctg aaggactttg tgagcgcttt gccagacaag ctcgaccatg aatgtgccga
     4381 aggcggtgag aacttaagcg taggccaacg tcagttagtt tgccttgctc gggcgctgct
     4441 ccgaaagact aagattctcg tattagatga agccactgcg gcagttgacc ttgaaacaga
     4501 tgacctcata cagagcacga tcagaacgca atttgaagac tgtacagttc tcaccatcgc
     4561 gcatagactt aacaccatta tggattacac aagagtcatt gtgttagaca agggtgagat
     4621 ccaagagtac ggggccccaa gtgatctgtt acaacaacga gggctgttct actctatggc
     4681 aaaggacgcc ggattagtat gacttgtgga aaggacgtgt cacttgaacg cgggacccag
     4741 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     4801 aggtcactat cagtcaaaat aaaatcatta tttgaggcta ggtggaggct cagtgatgat
     4861 aagtctgcga tggtggatgc atgtgtcatg gtcatagctg tttcctgtgt gaaattgtta
     4921 tccgctcaga gggcacaatc ctattccgcg ctatccgaca atctccaaga cattaggtgg
     4981 agttcagttc ggcgtatggc atatgtcgct ggaaagaaca tgtgagcaaa aggccagcaa
     5041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccacccccct
     5101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5701 gctctattca acaaagccgc cgtcccgtca agtcagcgta aatgggtagg gggcttcaaa
     5761 tcgtccgctc tgccagtgtt acaaccaatt aacaaattct gattagaaaa actcatcgag
     5821 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     5881 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     5941 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     6001 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     6061 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     6121 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     6181 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     6241 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     6301 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     6361 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     6421 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     6481 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     6541 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc aagacgtttc
     6601 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     6661 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     6721 gtggctttcc cccgccgctc tagaactagt ggatccaaat aaaacgaaag gctcagtcga
     6781 aagactgggc ctttcgtttt atctgttgtt tgtcgcatta tacgagacgt ccaggttggg
     6841 atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac
     6901 aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat gcataatccg
     6961 cacgcatctg gaataaggaa gtgccattcc gcctgacct
//
