LOCUS       sequence-10052-e        3428 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 3428)
  AUTHORS   Maeder ML, Thibodeau-Beganny S, Sander JD, Voytas DF, Joung JK
  TITLE     Oligomerized pool engineering (OPEN): an 'open-source' protocol for 
            making customized zinc-finger arrays.
  JOURNAL   Nat Protoc. 2009. 4(10):1471-501.
  PUBMED    19798082
REFERENCE   2  (bases 1 to 3428)
  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.
            2009. 4(10):1471-501."
COMMENT     Sequence Label: pKJ1267 (aka pAC-alphaGal4)
FEATURES             Location/Qualifiers
     source          1..3428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /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(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1624..1629,1630..1647,1648..1654)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1662..1678
                     /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     1667..1689
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1686..1702
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1686..1702
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(join(2988..3428,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagcttg catgcatcca tcaaaaaaat attgacaaca
     1561 taaaaaactt tgtgttatac ttgtaacgct acatggagat taactcaatc tagctagaga
     1621 ggctttacac tttatgcttc cggctcgtat aatgtgtgga attgtgagcg gataacaatt
     1681 tcacacagga aacagctatg accatgatta cggattcact ggaactctag accaaagaga
     1741 ggacacaatg cagggttctg tgacagagtt tctaaaaccg cgcctggttg atatcgagca
     1801 agtgagttcg acgcacgcca aggtgaccct tgagccttta gagcgtggct ttggccatac
     1861 tctgggtaac gcactgcgcc gtattctgct ctcatcgatg ccgggttgcg cggtgaccga
     1921 ggttgagatt gatggtgtac tacatgagta cagcaccaaa gaaggcgttc aggaagatat
     1981 cctggaaatc ctgctcaacc tgaaagggct ggcggtgaga gttcagggca aagatgaagt
     2041 tattcttacc ttgaataaat ctggcattgg ccctgtgact gcagccgata tcacccacga
     2101 cggtgatgtc gaaatcgtca agccgcagca cgtgatctgc cacctgaccg atgagaacgc
     2161 gtctattagc atgcgtatca aagttcagcg cggtcgtggt tatgtgccgg cttctacccg
     2221 aattcattcg gaagaagatg agcgcccaat cggccgtctg ctggtcgacg catgctacag
     2281 ccctgtggag cgtattgcct acaatgttga agcagcgcgt gtagaacagc gtaccgacct
     2341 ggacaagctg gtcatcgaaa tggaaaccaa cggcacaatc gatcctgaag aggcgattcg
     2401 tcgtgcggca accattctgg ctgaacaact ggaagctttc gttgacttac gtgatgtacg
     2461 tcagcctgaa gtgaaagaag agaaaccaga ggcggccgca gaatcaaggc tagaaagact
     2521 ggaacagcta tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga
     2581 ttctttacag gatataaaag cattgttaac aggattattt tgaagatctg catcgcagga
     2641 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgcta accgttttta
     2701 tcaggctctg ggaggcagaa taaatgatca tatcgtcaat tattacctcc acggggagag
     2761 cctgagcaaa ctggcctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     2821 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     2881 cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca
     2941 ggcgtagcac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     3001 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     3061 cagacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     3121 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     3181 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     3241 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     3301 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     3361 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     3421 gccatacg
//
