LOCUS       sequence-71992-e        5306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FX cloning Xenopus oocyte expression vector with SP6 promoter and 
            N-terminal chicken alpha7 signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5306)
  AUTHORS   Geertsma ER, Dutzler R
  TITLE     A versatile and efficient high-throughput cloning tool for 
            structural biology.
  JOURNAL   Biochemistry. 2011 Apr 19;50(15):3272-8. Epub 2011 Mar 25.
  PUBMED    21410291
REFERENCE   2  (bases 1 to 5306)
  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: 
            "Biochemistry."; date: "2011-04-19"; volume: "50(15)"; pages: 
            "3272-8. Epub 2011 Mar 25"
COMMENT     Sequence Label: pTLNXA7
FEATURES             Location/Qualifiers
     source          1..5306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(112..417)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(214..233)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(759..1418)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1333..1352
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1472..1478,1479..1496,1497..1502))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1527..1651)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1670..1690)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1694..1714)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1928..1944
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1929..1945
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     3'UTR           1995..2120
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2223..2239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2223..2239)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2236..2258)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2247..2263
                     /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)."
     promoter        complement(join(2271..2277,2278..2295,2296..2301))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2488..2505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2659..3247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2739..2758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3418..4209,4210..4278))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4041..4060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4279..4370)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4389..4416)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4552..4574)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4908..4927
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5218..5236
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5218..5235
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     5'UTR           5248..5290
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      join(5301..5306,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgggactga gagctctgat gctgtggctg ctggctgctg ctggactggt gagagaatct
       61 ctgagttgaa gagcgacctg cagactggct gtgtataagg gagcctgaca tttatattcc
      121 ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac
      181 ttcttccccg ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct
      241 ttcatccccg atatgcacca ccgggtaaag ttcacgggag actttatctg acagcagacg
      301 tgcactggcc agggggatca ccatccgtcg cccgggcgtg tcaataatat cactctgtac
      361 atccacaaac agacgataac ggctctctct tttataggtg taaaccttaa actgcatttc
      421 accagcccct gttctcgtca gcaaaagagc cgttcatttc aataaaccgg gcgacctcag
      481 ccatcccttc ctgattttcc gctttccagc gttcggcacg cagacgacgg gcttcattct
      541 gcatggttgt gcttaccaga ccggagatat tgacatcata tatgccttga gcaactgata
      601 gctgtcgctg tcaactgtca ctgtaatacg ctgcttcata gcatacctct ttttgacata
      661 cttcgggtat acatatcagt atatattctt ataccgcaaa aatcagcgcg caaatacgca
      721 tactgttatc tggcttttag taagccggat ccacgcgttt acgccccgcc ctgccactca
      781 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
      841 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
      901 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
      961 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     1021 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     1081 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     1141 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     1201 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     1261 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     1321 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     1381 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     1441 aatctcgacg gatcctaact caaaatccac acattatacg agccggaagc ataaagtgta
     1501 aagcctgggg tgcctaatgc ggccgccata gtgactggat atgttgtgtt ttacagtatt
     1561 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     1621 tttctcgttc agcttttttg tacaaacttg tgatatcgaa ttcgctagct acctttctct
     1681 tcttttttgg atctaccttt ctcttctttt ttggggatcc actagttcag ctcttctgca
     1741 taatccggtc tgaaggaggt ggctcgtctc gggggctggg gatgggatgg tgggtgttgg
     1801 atgcatctcc cgggatttat cttgagaata taaagaccca gaggttgaga tagagagaaa
     1861 gaaagatcgg gtcgaccacg aattcttgct ccaggtccag acaggacgct ttagagcaca
     1921 atggatctcg aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga
     1981 tctggttacc actaaaccag cctcaagaac acccgaatgg agtctctaag ctacataata
     2041 ccaacttaca ctttacaaaa tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa
     2101 taaaaagaaa gtttcttcac attctaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaagc
     2161 gcgcgcgcgc gcgcgcgctg gcggaccgac gcgttaacta cgtatttaaa ttcgtaatca
     2221 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2281 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctggcc cgtccggcca
     2341 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     2401 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggggcactt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3061 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3181 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3241 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3301 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3361 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3541 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3721 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4021 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4261 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa agagtttgta
     4381 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
     4441 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
     4501 ccggcctcag tggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4561 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4621 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     4681 agagcagatt gtactgagag tgcaccatat cgacgctctc ccttatgcga ctcctgcatt
     4741 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     4801 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     4861 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     4921 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     4981 tagaggatct ggctagcgat gaccctgctg attggttcgc tgaccatttc cggggtgcgg
     5041 aacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga cggcagttta
     5101 cgagagagat gatagggtct gcttcagtaa gccagatgct acacaattag gcttgtacat
     5161 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     5221 taggtgacac tatagaatac aagcttgctt gttctttttg cagaagctca gaataaacgc
     5281 tcaactttgg cagatcgcga gccacc
//
