LOCUS       sequence-308248-        4968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Cloning compatible entry vector for the human ACE2 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  AUTHORS   Chavez Lab
  TITLE     Coronavirus host receptors and proteases
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4968)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDONR221-ACE2
FEATURES             Location/Qualifiers
     source          1..4968
                     /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(3093..3192)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3209..3228)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3210..3228)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3351..3370)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3362..4171
                     /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(3429..3448)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4318..4906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4807..4826
                     /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctac aaaatgtcaa gctcttcctg gctccttctc agccttgttg ctgtaactgc
      721 tgctcagtcc accattgagg aacaggccaa gacatttttg gacaagttta accacgaagc
      781 cgaagacctg ttctatcaaa gttcacttgc ttcttggaat tataacacca atattactga
      841 agagaatgtc caaaacatga ataatgctgg ggacaaatgg tctgcctttt taaaggaaca
      901 gtccacactt gcccaaatgt atccactaca agaaattcag aatctcacag tcaagcttca
      961 gctgcaggct cttcagcaaa atgggtcttc agtgctctca gaagacaaga gcaaacggtt
     1021 gaacacaatt ctaaatacaa tgagcaccat ctacagtact ggaaaagttt gtaacccaga
     1081 taatccacaa gaatgcttat tacttgaacc aggtttgaat gaaataatgg caaacagttt
     1141 agactacaat gagaggctct gggcttggga aagctggaga tctgaggtcg gcaagcagct
     1201 gaggccatta tatgaagagt atgtggtctt gaaaaatgag atggcaagag caaatcatta
     1261 tgaggactat ggggattatt ggagaggaga ctatgaagta aatggggtag atggctatga
     1321 ctacagccgc ggccagttga ttgaagatgt ggaacatacc tttgaagaga ttaaaccatt
     1381 atatgaacat cttcatgcct atgtgagggc aaagttgatg aatgcctatc cttcctatat
     1441 cagtccaatt ggatgcctcc ctgctcattt gcttggtgat atgtggggta gattttggac
     1501 aaatctgtac tctttgacag ttccctttgg acagaaacca aacatagatg ttactgatgc
     1561 aatggtggac caggcctggg atgcacagag aatattcaag gaggccgaga agttctttgt
     1621 atctgttggt cttcctaata tgactcaagg attctgggaa aattccatgc taacggaccc
     1681 aggaaatgtt cagaaagcag tctgccatcc cacagcttgg gacctgggga agggcgactt
     1741 caggatcctt atgtgcacaa aggtgacaat ggacgacttc ctgacagctc atcatgagat
     1801 ggggcatatc cagtatgata tggcatatgc tgcacaacct tttctgctaa gaaatggagc
     1861 taatgaagga ttccatgaag ctgttgggga aatcatgtca ctttctgcag ccacacctaa
     1921 gcatttaaaa tccattggtc ttctgtcacc cgattttcaa gaagacaatg aaacagaaat
     1981 aaacttcctg ctcaaacaag cactcacgat tgttgggact ctgccattta cttacatgtt
     2041 agagaagtgg aggtggatgg tctttaaagg ggaaattccc aaagaccagt ggatgaaaaa
     2101 gtggtgggag atgaagcgag agatagttgg ggtggtggaa cctgtgcccc atgatgaaac
     2161 atactgtgac cccgcatctc tgttccatgt ttctaatgat tactcattca ttcgatatta
     2221 cacaaggacc ctttaccaat tccagtttca agaagcactt tgtcaagcag ctaaacatga
     2281 aggccctctg cacaaatgtg acatctcaaa ctctacagaa gctggacaga aactgttcaa
     2341 tatgctgagg cttggaaaat cagaaccctg gaccctagca ttggaaaatg ttgtaggagc
     2401 aaagaacatg aatgtaaggc cactgctcaa ctactttgag cccttattta cctggctgaa
     2461 agaccagaac aagaattctt ttgtgggatg gagtaccgac tggagtccat atgcagacca
     2521 aagcatcaaa gtgaggataa gcctaaaatc agctcttgga gataaagcat atgaatggaa
     2581 cgacaatgaa atgtacctgt tccgatcatc tgttgcatat gctatgaggc agtacttttt
     2641 aaaagtaaaa aatcagatga ttctttttgg ggaggaggat gtgcgagtgg ctaatttgaa
     2701 accaagaatc tcctttaatt tctttgtcac tgcacctaaa aatgtgtctg atatcattcc
     2761 tagaactgaa gttgaaaagg ccatcaggat gtcccggagc cgtatcaatg atgctttccg
     2821 tctgaatgac aacagcctag agtttctggg gatacagcca acacttggac ctcctaacca
     2881 gccccctgtt tccatatggc tgattgtttt tggagttgtg atgggagtga tagtggttgg
     2941 cattgtcatc ctgatcttca ctgggatcag agatcggaag aagaaaaata aagcaagaag
     3001 tggagaaaat ccttatgcct ccatcgatat tagcaaagga gaaaataatc caggattcca
     3061 aaacactgat gatgttcaga cctcctttta gaacccagct ttcttgtaca aagttggcat
     3121 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     3181 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     3241 tgtttcttgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     3301 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     3361 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     3421 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3481 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3541 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     3601 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3661 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3721 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3781 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3841 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3901 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3961 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     4021 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     4081 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     4141 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     4201 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     4261 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4321 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4381 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4441 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4501 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4561 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4621 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4681 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4741 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4801 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4861 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4921 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
