LOCUS       sequence-85608-e        6126 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 6126)
  AUTHORS   Gradia SD, Ishida JP, Tsai MS, Jeans C, Tainer JA, Fuss JO
  TITLE     MacroBac: New Technologies for Robust and Efficient Large-Scale 
            Production of Recombinant Multiprotein Complexes.
  JOURNAL   Methods Enzymol. 2017;592:1-26. doi: 10.1016/bs.mie.2017.03.008. 
            Epub 2017 May 15.
  PUBMED    28668116
REFERENCE   2  (bases 1 to 6126)
  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: "Methods 
            Enzymol."; date: "2017"; volume: "592:1-26. doi"; pages: " 
            10.1016/bs.mie.2017.03.008. Epub 2017 May 15"
COMMENT     Sequence Label: pFastBac TEV SNAPf Prescission TwinStrepII clo...
FEATURES             Location/Qualifiers
     source          1..6126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  68..292
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(359..892)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1062..1067,1068..1084,1085..1090))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     1499..1516
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2217..2240
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2250..2273
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2310..2333
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2621..2640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2675..2694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2792..2957
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3141..3596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3228..3247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3438..3459
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3623..3727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3728..3796,3797..4588)
                     /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(3946..3965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4759..5347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5248..5267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5501..5518
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5602..5621)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5762..5784
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5778..5969)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaccagccgc gtaacctggc aaaatcggtt acggttgagt aataaatgga tgccctgcgt
       61 aagcgggtgt gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca
      121 ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa
      181 aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt
      241 gcccgtcgta ttaaagaggg gcgtggccaa gggcatggta aagactatat tcgcggcgtt
      301 gtgacaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt
      361 aggtggcggt acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg
      421 tatagagagc cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac
      481 caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc
      541 ggtgctcgcc ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact
      601 tgggcagaac gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg
      661 cgatgaatgt cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg
      721 agtaggtggc tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt
      781 tgacttggtc agggccgagc ctacatgtgc gaatgatgcc catacttgag ccacctaact
      841 ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctccataac atcaaacatc
      901 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac
      961 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
     1021 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
     1081 cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
     1141 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
     1201 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
     1261 cggatctgcc ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg
     1321 tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt
     1381 tcgcccagga ctctagctat agttctagtg gttggctacg tatactccgg actattaata
     1441 gatcatggag ataattagcg gccgcgttta aacatccacc atttgatgga gataattaaa
     1501 atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa
     1561 aaaaacctat aactattccg gattattcat accgtcccac catcgggcgc ggatcccttt
     1621 aagaaggaga tatagttaac ctctacttcc aatccggctc tgataaagac tgcgagatga
     1681 agcgcacgac gctcgattcg ccccttggca aattggaact gagcggttgt gagcaaggtc
     1741 tgcatcgtat catctttctg ggaaagggaa ctagtgcagc agatgcagtc gaggtcccgg
     1801 ctcctgctgc cgtgcttggt ggtcccgagc ctttgatgca agctaccgct tggctgaatg
     1861 catatttcca tcagccagag gcgatcgagg agttccccgt tcccgcgctc catcatccgg
     1921 ttttccaaca ggaatccttc acccgccagg tcctgtggaa gctcctgaag gtcgtaaagt
     1981 ttggtgaagt gatctcatac agtcacttgg ccgccctggc gggcaaccca gccgctactg
     2041 ctgccgttaa gaccgcactc agtggcaacc ctgtacctat cctgatccct tgtcatagag
     2101 tcgtccaagg cgacctggac gtgggaggtt acgagggcgg tctggctgtg aaagaatggc
     2161 tgttggcaca tgagggccat cgtctgggca aacctggcct tggacccgcc ggtggtttgg
     2221 aggtactgtt ccagggcccg ggagcttcat ggtctcaccc ccaatttgag aaaggcggtg
     2281 gcagcggcgg cggtagcgga ggtggcagct ggtcacaccc acaattcgag aaataacggt
     2341 ccgaagcgcg cggaattcaa aggcctacgt cgacgagctc actagtcgcg gcctcatgat
     2401 ctcctagggg ccgctttcga atctagagcc tgcagtctcg aggcatgcgg taccaagctt
     2461 gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     2521 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2581 gtttacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2641 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2701 tcttatcatg tctggatctg atcactgctt gagcctagaa acatccacca tttaaatgga
     2761 gttaggttta aacgcctgca ggggagatcc gaaccagata agtgaaatct agttccaaac
     2821 tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt
     2881 ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac
     2941 tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat caaacatcct
     3001 gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac agaatgaaaa
     3061 tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa cgcttatttg
     3121 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3181 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3241 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3301 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3361 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3421 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3481 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3541 gctgatttaa caaaaattta acgcgaattt taacaaacta ttaacgttta caatttcagg
     3601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaatact attgaaaaag
     3721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3961 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4861 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5041 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5101 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5161 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5221 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5281 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5341 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5401 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5461 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5521 agcggaagag gcgcgccatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5581 gcatatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     5641 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     5701 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5761 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     5821 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     5881 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     5941 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     6001 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     6061 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     6121 gaccag
//
