LOCUS       sequence-85603-e        5446 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5446)
  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 5446)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 cloning vector with BioBrick PolyProm...
FEATURES             Location/Qualifiers
     source          1..5446
                     /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"
     polyA_signal    1904..2038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1941..1960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1995..2014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2112..2277
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2461..2916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2548..2567)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2758..2779
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2943..3047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3048..3116,3117..3908)
                     /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(3266..3285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4079..4667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4568..4587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4821..4838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4922..4941)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5082..5104
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5098..5289)
                     /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 ggatccttta
     1621 ttaagtactt ccaatccaat attggaagtg gataaccggt ccgaagcgcg cggaattcaa
     1681 aggcctacgt cgacgagctc actagtcgcg gcctcatgat ctcctagggg ccgctttcga
     1741 atctagagcc tgcagtctcg aggcatgcgg taccaagctt gtcgagaagt actagaggat
     1801 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1861 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1921 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1981 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg
     2041 atcactgctt gagcctagaa acatccacca tttaaatgga gttaggttta aacgcctgca
     2101 ggggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt
     2161 tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata
     2221 atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc gcccacagcg
     2281 gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca tgtgacaaac
     2341 cgtcatcttc ggctactttt tctctgtcac agaatgaaaa tttttctgtc atctcttcgt
     2401 tattaatgtt tgtaattgac tgaatatcaa cgcttatttg cagcctgaat ggcgaatggg
     2461 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2521 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2581 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2641 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2701 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2761 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2821 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2881 acgcgaattt taacaaacta ttaacgttta caatttcagg tggcactttt cggggaaatg
     2941 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3001 gacaataacc ctgataaatg cttcaatact attgaaaaag gaagagtatg agtattcaac
     3061 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3121 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3181 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3241 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3301 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3361 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3421 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3481 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3541 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3601 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3661 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3721 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3781 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3841 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3901 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3961 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4021 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4081 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4141 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4201 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4261 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4321 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4381 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4441 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4501 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4561 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4621 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4681 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4741 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4801 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag gcgcgccatg
     4861 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca
     4921 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     4981 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5041 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5101 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5161 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5221 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5281 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     5341 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     5401 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccag
//
