LOCUS       sequence-71990-e        5246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FX cloning Xenopus oocyte expression vector with SP6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5246)
  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 5246)
  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: pTLNX
FEATURES             Location/Qualifiers
     source          1..5246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(52..357)
                     /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(154..173)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(699..1358)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1273..1292
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1412..1418,1419..1436,1437..1442))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1467..1591)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1610..1630)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1634..1654)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1868..1884
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1869..1885
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     3'UTR           1935..2060
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2163..2179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2163..2179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2176..2198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2187..2203
                     /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(2211..2217,2218..2235,2236..2241))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2428..2445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2599..3187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2679..2698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3358..4149,4150..4218))
                     /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     3981..4000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4219..4310)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4329..4356)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4492..4514)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4848..4867
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5158..5176
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5158..5175
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atgagttgaa gagcgacctg cagactggct gtgtataagg gagcctgaca tttatattcc
       61 ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac
      121 ttcttccccg ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct
      181 ttcatccccg atatgcacca ccgggtaaag ttcacgggag actttatctg acagcagacg
      241 tgcactggcc agggggatca ccatccgtcg cccgggcgtg tcaataatat cactctgtac
      301 atccacaaac agacgataac ggctctctct tttataggtg taaaccttaa actgcatttc
      361 accagcccct gttctcgtca gcaaaagagc cgttcatttc aataaaccgg gcgacctcag
      421 ccatcccttc ctgattttcc gctttccagc gttcggcacg cagacgacgg gcttcattct
      481 gcatggttgt gcttaccaga ccggagatat tgacatcata tatgccttga gcaactgata
      541 gctgtcgctg tcaactgtca ctgtaatacg ctgcttcata gcatacctct ttttgacata
      601 cttcgggtat acatatcagt atatattctt ataccgcaaa aatcagcgcg caaatacgca
      661 tactgttatc tggcttttag taagccggat ccacgcgttt acgccccgcc ctgccactca
      721 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
      781 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
      841 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
      901 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
      961 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     1021 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     1081 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     1141 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     1201 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     1261 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     1321 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     1381 aatctcgacg gatcctaact caaaatccac acattatacg agccggaagc ataaagtgta
     1441 aagcctgggg tgcctaatgc ggccgccata gtgactggat atgttgtgtt ttacagtatt
     1501 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     1561 tttctcgttc agcttttttg tacaaacttg tgatatcgaa ttcgctagct acctttctct
     1621 tcttttttgg atctaccttt ctcttctttt ttggggatcc actagttcag ctcttctgca
     1681 taatccggtc tgaaggaggt ggctcgtctc gggggctggg gatgggatgg tgggtgttgg
     1741 atgcatctcc cgggatttat cttgagaata taaagaccca gaggttgaga tagagagaaa
     1801 gaaagatcgg gtcgaccacg aattcttgct ccaggtccag acaggacgct ttagagcaca
     1861 atggatctcg aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga
     1921 tctggttacc actaaaccag cctcaagaac acccgaatgg agtctctaag ctacataata
     1981 ccaacttaca ctttacaaaa tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa
     2041 taaaaagaaa gtttcttcac attctaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaagc
     2101 gcgcgcgcgc gcgcgcgctg gcggaccgac gcgttaacta cgtatttaaa ttcgtaatca
     2161 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2221 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctggcc cgtccggcca
     2281 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     2341 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2401 ataccgcatc aggggcactt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2461 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2521 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2581 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2641 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2701 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2761 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2821 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2881 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2941 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3001 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3061 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3121 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3181 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3301 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3361 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3421 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3481 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3541 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3601 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3661 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3721 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3781 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3841 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3901 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3961 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4021 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4081 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4141 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4201 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa agagtttgta
     4321 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
     4381 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
     4441 ccggcctcag tggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4501 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4561 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     4621 agagcagatt gtactgagag tgcaccatat cgacgctctc ccttatgcga ctcctgcatt
     4681 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     4741 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     4801 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     4861 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     4921 tagaggatct ggctagcgat gaccctgctg attggttcgc tgaccatttc cggggtgcgg
     4981 aacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga cggcagttta
     5041 cgagagagat gatagggtct gcttcagtaa gccagatgct acacaattag gcttgtacat
     5101 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     5161 taggtgacac tatagaatac aagcttgctt gttctttttg cagaagctca gaataaacgc
     5221 tcaactttag cagatcgcga gccacc
//
