LOCUS       sequence-85606-e        6202 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 6202)
  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 6202)
  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 StrepII msfGFP TEV cloning vector wit...
FEATURES             Location/Qualifiers
     source          1..6202
                     /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             1633..1656
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             join(1672..1674,1675..2385)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1714..1735)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2322..2343
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2395..2415
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    2660..2794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2697..2716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2751..2770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2868..3033
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3217..3672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3304..3323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3514..3535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3699..3803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3804..3872,3873..4664)
                     /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(4022..4041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4835..5423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5324..5343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5577..5594
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5678..5697)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5838..5860
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5854..6045)
                     /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 ggatcccggt
     1621 ccgatggcta gctggagcca cccgcagttc gaaaaaggcg ccggttcttc tgtgagcaag
     1681 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1741 ggccacaagt tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc
     1801 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1861 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1921 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatctcctt caaggacgac
     1981 ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2041 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2101 aacttcaaca gccacaacgt ctatatcacg gccgacaagc agaagaacgg catcaaggcg
     2161 aacttcaaga tccgccacaa cgtcgaggac ggcagcgtgc agctcgccga ccactaccag
     2221 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2281 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2341 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaaggggat cgaggaaaac
     2401 ctgtacttcc aatccaatat tggaagtgga taacggatcc gaattcgagc gccgtcgaca
     2461 agcttgcggc ctcatgatct cctaggggcc tcatgatctc ctaggggccg cactcgaggc
     2521 atgcggtacc aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt
     2581 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2641 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2701 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2761 caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc ctagaaacat
     2821 ccaccattta aatggagtta ggtttaaacg cctgcagggg agatccgaac cagataagtg
     2881 aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac
     2941 ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc
     3001 cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt
     3061 tttaatcaaa catcctgcca actccatgtg acaaaccgtc atcttcggct actttttctc
     3121 tgtcacagaa tgaaaatttt tctgtcatct cttcgttatt aatgtttgta attgactgaa
     3181 tatcaacgct tatttgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3241 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3301 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3361 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3421 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3481 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3541 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3601 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaactattaa
     3661 cgtttacaat ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3721 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3781 aatactattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3841 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3901 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3961 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4021 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4081 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4141 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4201 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4261 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4321 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4381 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4441 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4501 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4561 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4621 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4681 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4741 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4801 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4861 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4921 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4981 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5041 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5101 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5161 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5221 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5281 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5341 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5401 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5461 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5521 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5581 agtcagtgag cgaggaagcg gaagaggcgc gccatgcggt attttctcct tacgcatctg
     5641 tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat
     5701 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     5761 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5821 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5881 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     5941 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     6001 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     6061 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     6121 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     6181 ggtatggatg cggcgggacc ag
//
