LOCUS       sequence-404029-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBIG2abc - Step2 vector abc (biGBac system for expression of protein
            complexes in insect cells).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5669)
  AUTHORS   Weissmann F, Petzold G, VanderLinden R, Huis In 't Veld PJ, Brown 
            NG, Lampert F, Westermann S, Stark H, Schulman BA, Peters JM
  TITLE     biGBac enables rapid gene assembly for the expression of large 
            multisubunit protein complexes.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2564-9. doi: 
            10.1073/pnas.1604935113. Epub 2016 Apr 25.
  PUBMED    27114506
REFERENCE   2  (bases 1 to 5669)
  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"; doi: 
            "10.1073/pnas.1604935113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-05-10- 10"; volume: "113"; issue: "19"; pages: "E2564-9"
COMMENT     Sequence Label: pBIG2abc
FEATURES             Location/Qualifiers
     source          1..5669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(339..344,345..361,362..367))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(763..796)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  874..1039
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        1424..1526
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1527..2186
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1593..1612)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2486..2940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2573..2592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2782..2803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2966..3070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3071..3139,3140..3931)
                     /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(3289..3308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4102..4690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4591..4610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4844..4861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4995..5219
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(5286..5669,1..150))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct
       61 acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg
      121 ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac
      181 ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt
      241 cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc
      301 tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt
      361 atgtcaactg ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg
      421 gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct
      481 ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg
      541 atctgccctt gcttcaggag atcggtagac ctcggccgtc gcggcgcttg ccggtggtgc
      601 tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg
      661 cccaggactc tagctatagt tctagtggtt ggctacagct ttgtttgtac tatcaacagg
      721 ttgaactgct gatcaacaga tcctctacgc ggccgcggta ccataacttc gtatagcata
      781 cattatacga agttatctgg cctaggttaa ttaaacgctc tatggtctaa agtttaaacc
      841 taatgatgcc tgatgtttca atcaggagat ccgaaccaga taagtgaaat ctagttccaa
      901 actattttgt catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta
      961 ttttgtcact cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca
     1021 actattttgt ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc
     1081 ctgccaactc catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa
     1141 aatttttctg tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt
     1201 tgcagcctga actgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
     1261 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
     1321 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     1381 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     1441 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     1501 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     1561 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     1621 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     1681 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     1741 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     1801 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     1861 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     1921 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     1981 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     2041 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     2101 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     2161 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
     2221 ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa
     2281 attcgacccg gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg
     2341 gtttaccggt ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg
     2401 ccagtttgct caggctctcc ccgtggaggt aataattgac gatatgatca tttattctgc
     2461 ctcccagagc ctgatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2521 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2641 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2701 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2761 ttggagtcca cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2821 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2881 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2941 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3001 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3061 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3121 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3181 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3241 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3301 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3361 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3421 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3481 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3541 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3601 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3661 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3721 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3781 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3841 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3901 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3961 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4021 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4081 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4141 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4201 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactattc ttctagtgta
     4261 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4321 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4381 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4441 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4501 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4561 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4621 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4681 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4741 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4801 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4861 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4921 ccgcatagac cagccgcgta acctggcaaa atcggttacg gttgagtaat aaatggatgc
     4981 cctgcgtaag cgggtgtggg cggacaataa agtcttaaac tgaacaaaat agatctaaac
     5041 tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag tccagttatg
     5101 ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat tttctgaagt
     5161 gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg catggtaaag actatattcg
     5221 cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg
     5281 aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc
     5341 aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca
     5401 taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc
     5461 tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc
     5521 tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca
     5581 gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga
     5641 tgttgggagt aggtggctac gtctccgaa
//
