LOCUS       sequence-85601-e        6073 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 6073)
  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 6073)
  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 TwinStrepII Prescission SNAPf TEV clo...
FEATURES             Location/Qualifiers
     source          1..6073
                     /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     1474..1491
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1601..1624
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1661..1684
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1685..1708
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2273..2293
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    2560..2694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2597..2616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2651..2670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2739..2904
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3088..3543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3175..3194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3385..3406
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3570..3674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3675..3743,3744..4535)
                     /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(3893..3912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4706..5294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5195..5214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5448..5465
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5549..5568)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5709..5731
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5725..5916)
                     /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 gccgcttaat taaaaatgat aaccatctcg caaataaata
     1501 agtattttac tgttttcgta acagttttgt aataaaaaaa cctataacta ttccggatta
     1561 ttcataccgt cccaccatcg ggcgcggatc catggcttca tggtctcacc cccaatttga
     1621 gaaaggcggt ggcagcggcg gcggtagcgg aggtggcagc tggtcacacc cacaattcga
     1681 gaaattggag gtactgttcc agggcccggg aagcggcgat aaagactgcg agatgaagcg
     1741 cacgacgctc gattcgcccc ttggcaaatt ggaactgagc ggttgtgagc aaggtctgca
     1801 tcgtatcatc tttctgggaa agggaactag tgcagcagat gcagtcgagg tcccggctcc
     1861 tgctgccgtg cttggtggtc ccgagccttt gatgcaagct accgcttggc tgaatgcata
     1921 tttccatcag ccagaggcga tcgaggagtt ccccgttccc gcgctccatc atccggtttt
     1981 ccaacaggaa tccttcaccc gccaggtcct gtggaagctc ctgaaggtcg taaagtttgg
     2041 tgaagtgatc tcatacagtc acttggccgc cctggcgggc aacccagccg ctactgctgc
     2101 cgttaagacc gcactcagtg gcaaccctgt acctatcctg atcccttgtc atagagtcgt
     2161 ccaaggcgac ctggacgtgg gaggttacga gggaggtctg gctgtgaaag aatggctgtt
     2221 ggcacatgag ggccatcgtc tgggcaaacc tggccttgga cccgccggtg gtgaaaacct
     2281 gtacttccaa tccaatattg gaagtggata accggtccga agcgcgcgga attcaaaggc
     2341 ctacgtcgac gagctcacta gtcgcggcct catgatctcc taggggccgc tttcgaatct
     2401 agagcctgca gtctcgaggc atgcggtacc aagcttgtcg agaagtacta gaggatcata
     2461 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2521 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca
     2701 ctgcttgagc ctagcgatcg ccctgcaggg gagatccgaa ccagataagt gaaatctagt
     2761 tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc
     2821 atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt
     2881 tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa
     2941 acatcctgcc aactccatgt gacaaaccgt catcttcggc tactttttct ctgtcacaga
     3001 atgaaaattt ttctgtcatc tcttcgttat taatgtttgt aattgactga atatcaacgc
     3061 ttatttgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     3121 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3181 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3241 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3301 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3361 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3421 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3481 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaactatta acgtttacaa
     3541 tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3601 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caatactatt
     3661 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3721 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3781 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3841 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3901 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3961 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4021 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4081 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4141 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4201 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4261 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4321 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4381 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4441 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4501 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4561 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4621 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4681 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4741 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4801 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4861 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4921 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4981 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5041 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5101 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5161 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5221 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5281 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5341 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5401 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5461 gcgaggaagc ggaagaggcg cgccatgcgg tattttctcc ttacgcatct gtgcggtatt
     5521 tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5581 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     5641 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5701 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5761 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5821 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5881 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     5941 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     6001 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     6061 gcggcgggac cag
//
