LOCUS       sequence-69779-e        6148 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 6148)
  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 6148)
  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..6148
                     /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             1608..1631
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             join(1647..1649,1650..2360)
                     /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(1689..1710)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2297..2318
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2370..2390
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    2635..2769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2672..2691)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2726..2745
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2814..2979
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3163..3618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3250..3269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3460..3481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3645..3749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3750..3818,3819..4610)
                     /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(3968..3987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4781..5369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5270..5289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5523..5540
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5624..5643)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5784..5806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5800..5991)
                     /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 ggctagctgg agccacccgc
     1621 agttcgaaaa aggcgccggt tcttctgtga gcaagggcga ggagctgttc accggggtgg
     1681 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgcgcggcg
     1741 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca
     1801 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1861 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1921 acgtccagga gcgcaccatc tccttcaagg acgacggcac ctacaagacc cgcgccgagg
     1981 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2041 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata
     2101 tcacggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacgtcg
     2161 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2221 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     2281 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2341 gcatggacga gctgtacaag gggatcgagg aaaacctgta cttccaatcc aatattggaa
     2401 gtggataacg gatccgaatt cgagcgccgt cgacaagctt gcggcctcat gatctcctag
     2461 gggcctcatg atctcctagg ggccgcactc gaggcatgcg gtaccaagct tgtcgagaag
     2521 tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2581 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2641 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2701 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2761 gtctggatct gatcactgct tgagcctagc gatcgccctg caggggagat ccgaaccaga
     2821 taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg
     2881 ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt
     2941 tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt
     3001 tatgttttta atcaaacatc ctgccaactc catgtgacaa accgtcatct tcggctactt
     3061 tttctctgtc acagaatgaa aatttttctg tcatctcttc gttattaatg tttgtaattg
     3121 actgaatatc aacgcttatt tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3181 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3241 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3301 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3361 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3421 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3481 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3541 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaac
     3601 tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3661 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3721 tgcttcaata ctattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3781 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3841 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3901 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3961 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4021 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4081 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4141 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4201 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4261 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4321 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4381 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4441 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4501 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4561 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4621 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4681 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4741 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4801 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4861 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4921 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4981 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5041 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5101 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5161 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5221 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5281 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5341 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5401 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5461 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5521 gcagcgagtc agtgagcgag gaagcggaag aggcgcgcca tgcggtattt tctccttacg
     5581 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     5641 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     5701 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5761 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5821 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5881 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5941 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     6001 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     6061 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     6121 actggcggta tggatgcggc gggaccag
//
