LOCUS       sequence-69777-e        5407 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 5407)
  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 5407)
  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 His6 TEV cloning vector with BioBrick...
FEATURES             Location/Qualifiers
     source          1..5407
                     /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             1611..1628
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1629..1649
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    1894..2028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1931..1950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1985..2004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2073..2238
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2422..2877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2509..2528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2719..2740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2904..3008
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3009..3077,3078..3869)
                     /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(3227..3246)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4040..4628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4529..4548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4782..4799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4883..4902)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5043..5065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5059..5250)
                     /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 ccggtccgat gggttcttct caccatcacc
     1621 atcaccatga aaacctgtac ttccaatcca atattggaag tggataacgg atccgaattc
     1681 gagcgccgtc gacaagcttg cggcctcatg atctcctagg ggcctcatga tctcctaggg
     1741 gccgcactcg aggcatgcgg taccaagctt gtcgagaagt actagaggat cataatcagc
     1801 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1861 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1921 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1981 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt
     2041 gagcctagcg atcgccctgc aggggagatc cgaaccagat aagtgaaatc tagttccaaa
     2101 ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat
     2161 tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa
     2221 ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc
     2281 tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa
     2341 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt
     2401 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2461 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2521 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2581 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2641 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2701 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2761 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2821 agctgattta acaaaaattt aacgcgaatt ttaacaaact attaacgttt acaatttcag
     2881 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2941 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaatac tattgaaaaa
     3001 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3061 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3121 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3181 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3241 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3301 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3361 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3421 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3481 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3541 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3601 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3661 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3721 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3781 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3841 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3901 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3961 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4021 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4081 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4141 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4201 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4261 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4321 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4381 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4441 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4501 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4561 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4621 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4681 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4741 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4801 aagcggaaga ggcgcgccat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4861 cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4921 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4981 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5041 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     5101 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     5161 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     5221 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     5281 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     5341 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     5401 ggaccag
//
