LOCUS       sequence-7622-ex        4506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4506)
  AUTHORS   Wright DA, Thibodeau-Beganny S, Sander JD, Winfrey RJ, Hirsh AS, 
            Eichtinger M, Fu F, Porteus MH, Dobbs D, Voytas DF, Joung JK
  TITLE     Standardized reagents and protocols for engineering zinc finger 
            nucleases by modular assembly.
  JOURNAL   Nat Protoc. 2006;1(3):1637-52. doi: 10.1038/nprot.2006.259
  PUBMED    17406455
REFERENCE   2  (bases 1 to 4506)
  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: "Nat 
            Protoc."; date: "2006"; volume: "1(3):1637-52. doi"; pages: " 
            10.1038/nprot.2006.25"
COMMENT     Sequence Label: pAC-Kan-alphaGal4
FEATURES             Location/Qualifiers
     source          1..4506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(595..1410)
                     /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(595..1410)
                     /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     1318..1337
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1402..1421
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1871..1888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2005..2550)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2085..2104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2729..2747
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2999..3004,3005..3022,3023..3029)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    3037..3053
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3042..3064
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3061..3077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3061..3077
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccatggctga tcctccggcg ttcagcctgt gccacagccg acaggatggt gaccaccatt
       61 tgccccatat caccgtcggt actgatcccg tcgtcaataa accgaaccgc tacaccctga
      121 gcatcaaact cttttatcag ttggatcatg tcggcggtgt cgcggccaag acggtcgagc
      181 ttcttcacca gaatgacatc accttcctcc accttcatcc tcagcaaatc cagcccttcc
      241 cgatctgttg aactgccgga tgccttgtcg gtaaagatgc ggttagcttt tacccctgca
      301 tctttgagcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa
      361 tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg
      421 tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa
      481 gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccgccgtc
      541 ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa
      601 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
      661 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
      721 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
      781 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
      841 cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt
      901 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
      961 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     1021 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     1081 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     1141 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     1201 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     1261 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     1321 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
     1381 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
     1441 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
     1501 ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat
     1561 cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa
     1621 ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga
     1681 tggggcgatt caggcctggt atgagtcagc aacaccttct tcacgaggca gacctcagcg
     1741 ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt
     1801 catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat
     1861 attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa
     1921 tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag
     1981 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
     2041 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2101 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
     2161 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
     2221 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
     2281 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
     2341 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
     2401 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
     2461 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
     2521 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat
     2581 ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     2641 tataagttgt aattctcatg tttgacagct tatcatcgat aagctggacg tctaagaaac
     2701 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttggataacc
     2761 agaagcaata aaaaatcaaa tcggatttca ctatataatc tcactttatc taagatgaat
     2821 ccgatggaag catcctgttt tctctcaatt tttttatcta aaacccagcg ttcgatgctt
     2881 ctttgagcga acgatcaaaa ataagtgcct tcccatcaaa aaaatattga caacataaaa
     2941 aactttgtgt tatacttgta acgctacatg gagattaact caatctagct agagaggctt
     3001 tacactttat gcttccggct cgtataatgt gtggaattgt gagcggataa caatttcaca
     3061 caggaaacag ctatgaccat gattacggat tcactggaac tctagaccaa agagaggaca
     3121 caatgcaggg ttctgtgaca gagtttctaa aaccgcgcct ggttgatatc gagcaagtga
     3181 gttcgacgca cgccaaggtg acccttgagc ctttagagcg tggctttggc catactctgg
     3241 gtaacgcact gcgccgtatt ctgctctcat cgatgccggg ttgcgcggtg accgaggttg
     3301 agattgatgg tgtactacat gagtacagca ccaaagaagg cgttcaggaa gatatcctgg
     3361 aaatcctgct caacctgaaa gggctggcgg tgagagttca gggcaaagat gaagttattc
     3421 ttaccttgaa taaatctggc attggccctg tgactgcagc cgatatcacc cacgacggtg
     3481 atgtcgaaat cgtcaagccg cagcacgtga tctgccacct gaccgatgag aacgcgtcta
     3541 ttagcatgcg tatcaaagtt cagcgcggtc gtggttatgt gccggcttct acccgaattc
     3601 attcggaaga agatgagcgc ccaatcggcc gtctgctggt cgacgcatgc tacagccctg
     3661 tggagcgtat tgcctacaat gttgaagcag cgcgtgtaga acagcgtacc gacctggaca
     3721 agctggtcat cgaaatggaa accaacggca caatcgatcc tgaagaggcg attcgtcgtg
     3781 cggcaaccat tctggctgaa caactggaag ctttcgttga cttacgtgat gtacgtcagc
     3841 ctgaagtgaa agaagagaaa ccagaggcgg ccgcagaatc aaggctagaa agactggaac
     3901 agctatttct actgattttt cctcgagaag accttgacat gattttgaaa atggattctt
     3961 tacaggatat aaaagcattg ttaacaggat tattttgacc taggatctgc atcgcaggat
     4021 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctaa ccgtttttat
     4081 caggctctgg gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc
     4141 ctgagcaaac tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     4201 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     4261 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     4321 gcgtagcacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     4381 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     4441 agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     4501 atttgc
//
