LOCUS       sequence-69776-e        5392 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 5392)
  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 5392)
  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 LIC cloning vector with BioBrick
            poly...
FEATURES             Location/Qualifiers
     source          1..5392
                     /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"
     polyA_signal    1879..2013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1916..1935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1970..1989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2058..2223
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2407..2862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2494..2513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2704..2725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2889..2993
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2994..3062,3063..3854)
                     /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(3212..3231)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4025..4613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4514..4533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4767..4784
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4868..4887)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5028..5050
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5044..5235)
                     /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 ctttattaag tacttccaat ccaatattgg
     1621 aagtggataa ccggtccgaa gcgcgcggaa ttcaaaggcc tacgtcgacg agctcactag
     1681 tcgcggcctc atgatctcct aggggccgct ttcgaatcta gagcctgcag tctcgaggca
     1741 tgcggtacca agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta
     1801 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1861 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1921 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1981 aaactcatca atgtatctta tcatgtctgg atctgatcac tgcttgagcc tagcgatcgc
     2041 cctgcagggg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt
     2101 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc
     2161 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc
     2221 acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg
     2281 acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct
     2341 cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg
     2401 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2461 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2521 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2581 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2641 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2701 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2761 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2821 aatttaacgc gaattttaac aaactattaa cgtttacaat ttcaggtggc acttttcggg
     2881 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2941 tcatgagaca ataaccctga taaatgcttc aatactattg aaaaaggaag agtatgagta
     3001 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3061 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3121 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3181 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3241 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3301 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3361 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3421 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3481 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3541 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3601 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3661 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3721 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3781 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3841 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3901 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3961 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4021 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4081 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4141 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4201 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4261 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4321 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4381 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4441 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4501 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4561 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4621 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4681 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4741 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagaggcgc
     4801 gccatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca
     4861 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     4921 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     4981 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5041 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     5101 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     5161 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     5221 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     5281 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     5341 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc ag
//
