LOCUS       sequence-72043-e        7720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FX cloning insect cell expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7720)
  AUTHORS   Weinert C, Morger D, Djekic A, Grutter MG, Mittl PR
  TITLE     Crystal structure of TRIM20 C-terminal coiled-coil/B30.2 fragment: 
            implications for the recognition of higher order oligomers.
  JOURNAL   Sci Rep. 2015 Jun 4;5:10819. doi: 10.1038/srep10819.
  PUBMED    26043233
REFERENCE   2  (bases 1 to 7720)
  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: "Sci Rep."; 
            date: "2015-06-4"; volume: "5:10819. doi"; pages: " 
            10.1038/srep10819"
COMMENT     Sequence Label: pFBXC3GH
FEATURES             Location/Qualifiers
     source          1..7720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4169..4217
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4242..4261
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4353..4462)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4507..4598
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4523..4540
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             complement(4743..5048)
                     /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(4845..4864)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(5390..6049)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5964..5983
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6103..6109,6110..6127,6128..6133))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(6158..6282)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6301..6321)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6325..6345)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6372..6395
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6402..7112
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(6429..6457)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7052..7072
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             7119..7148
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    7310..7444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7347..7366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7401..7420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7473..7638
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggactagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cagctttgtt taaacaaagc tggctatggc agggcttgcc
     4021 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttgggggttg gggtggggaa
     4081 aaggaagaaa cgcgggcgta ttggtcccaa tggggtctcg gtggggtatc gacagagtgc
     4141 cagccctggg accgaacccc gcgtttatga acaaacgacc caacacccgt gcgttttatt
     4201 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     4261 agttagcctc ccccatctcc cggtaccgca tgctatgcat cagctgctag caccatggct
     4321 cgagatcccg ggtgatcaag tcttcgtcga gtgattgtaa ataaaatgta atttacagta
     4381 tagtatttta attaatatac aaatgatttg ataataattc ttatttaact ataatatatt
     4441 gtgttgggtt gaattaaagg tccgtatact agtatcgatt cgcgacctac tccggaatat
     4501 taatagatca tggagataat taaaatgata accatctcgc aaataaataa gtattttact
     4561 gttttcgtaa cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc
     4621 ccaccatcgg gcgcggatcc cggtccgaag cgcgcggaat tcaaaggcct acgtcgacga
     4681 gctcatacac catgagttga agagcgacct gcagactggc tgtgtataag ggagcctgac
     4741 atttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg
     4801 agatcagcca cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc
     4861 atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct
     4921 gacagcagac gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata
     4981 tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta
     5041 aactgcattt caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg
     5101 ggcgacctca gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg
     5161 ggcttcattc tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg
     5221 agcaactgat agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc
     5281 tttttgacat acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc
     5341 gcaaatacgc atactgttat ctggctttta gtaagccgga tccacgcgtt tacgccccgc
     5401 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     5461 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     5521 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     5581 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     5641 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     5701 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     5761 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     5821 ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     5881 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     5941 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     6001 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     6061 tagctcctga aaatctcgac ggatcctaac tcaaaatcca cacattatac gagccggaag
     6121 cataaagtgt aaagcctggg gtgcctaatg cggccgccat agtgactgga tatgttgtgt
     6181 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     6241 atcattttac gtttctcgtt cagctttttt gtacaaactt gtgatatcga attcgctagc
     6301 tacctttctc ttcttttttg gatctacctt tctcttcttt tttggggatc cactagttca
     6361 gctcttctgc attagaagtt ttgtttcaag gtccacaatt cagcaaagga gaagaacttt
     6421 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     6481 ctgtcagtgg agagggtgaa ggtgatgcta catacggaaa actcaccctt aaatttattt
     6541 gcactactgg aaaactacct gttccttggc caacacttgt cactactctg acctatggtg
     6601 ttcaatgctt ttcccgttat ccggatcaca tgaaacggca tgactttttc aagagtgcca
     6661 tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg aactacaaga
     6721 cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag ttaaagggta
     6781 ttgattttaa agaagatgga aacattctcg gacacaaact agagtacaac tataactcac
     6841 acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaattc
     6901 gccacaacat tgaagatggt tccgttcaac tagcagacca ttatcaacaa aatactccaa
     6961 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa tctgcccttt
     7021 cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta actgctgctg
     7081 ggattacaca tggcatggat gagctctaca aaactagtca tcatcatcac catcatcatc
     7141 accatcatta agtcgaccat ctagagcctg cagtctcgac aagcttgtcg agaagtacta
     7201 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     7261 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     7321 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7381 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7441 gatctgatca ctgcttgagc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa
     7501 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     7561 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     7621 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc
     7681 tgccaactcc atgtgacaaa ccgtcatctt cggctacttt
//
