LOCUS       sequence-85607-e        6127 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 6127)
  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 6127)
  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..6127
                     /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             1626..1649
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1686..1709
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1710..1733
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2298..2318
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    2585..2719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2622..2641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2676..2695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2793..2958
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3142..3597
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3229..3248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3439..3460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3624..3728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3729..3797,3798..4589)
                     /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(3947..3966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4760..5348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5249..5268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5502..5519
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5603..5622)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5763..5785
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5779..5970)
                     /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 ggatccatgg
     1621 cttcatggtc tcacccccaa tttgagaaag gcggtggcag cggcggcggt agcggaggtg
     1681 gcagctggtc acacccacaa ttcgagaaat tggaggtact gttccagggc ccgggaagcg
     1741 gcgataaaga ctgcgagatg aagcgcacga cgctcgattc gccccttggc aaattggaac
     1801 tgagcggttg tgagcaaggt ctgcatcgta tcatctttct gggaaaggga actagtgcag
     1861 cagatgcagt cgaggtcccg gctcctgctg ccgtgcttgg tggtcccgag cctttgatgc
     1921 aagctaccgc ttggctgaat gcatatttcc atcagccaga ggcgatcgag gagttccccg
     1981 ttcccgcgct ccatcatccg gttttccaac aggaatcctt cacccgccag gtcctgtgga
     2041 agctcctgaa ggtcgtaaag tttggtgaag tgatctcata cagtcacttg gccgccctgg
     2101 cgggcaaccc agccgctact gctgccgtta agaccgcact cagtggcaac cctgtaccta
     2161 tcctgatccc ttgtcataga gtcgtccaag gcgacctgga cgtgggaggt tacgagggag
     2221 gtctggctgt gaaagaatgg ctgttggcac atgagggcca tcgtctgggc aaacctggcc
     2281 ttggacccgc cggtggtgaa aacctgtact tccaatccaa tattggaagt ggataaccgg
     2341 tccgaagcgc gcggaattca aaggcctacg tcgacgagct cactagtcgc ggcctcatga
     2401 tctcctaggg gccgctttcg aatctagagc ctgcagtctc gaggcatgcg gtaccaagct
     2461 tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2521 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2581 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2641 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2701 atcttatcat gtctggatct gatcactgct tgagcctaga aacatccacc atttaaatgg
     2761 agttaggttt aaacgcctgc aggggagatc cgaaccagat aagtgaaatc tagttccaaa
     2821 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     2881 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     2941 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc
     3001 tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa
     3061 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt
     3121 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3181 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3241 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3301 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3361 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3421 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3481 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3541 agctgattta acaaaaattt aacgcgaatt ttaacaaact attaacgttt acaatttcag
     3601 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3661 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaatac tattgaaaaa
     3721 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3781 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3841 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3901 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3961 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4021 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4081 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4141 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4201 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4261 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4321 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4381 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4441 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4501 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4561 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4741 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4801 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4861 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4921 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4981 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5041 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5101 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5161 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5221 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5281 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5341 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5401 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5461 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5521 aagcggaaga ggcgcgccat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5581 cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     5641 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     5701 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5761 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     5821 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     5881 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     5941 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     6001 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     6061 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     6121 ggaccag
//
