LOCUS       sequence-354512-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBIG2ab - Step2 vector ab (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: pBIG2ab
FEATURES             Location/Qualifiers
     source          1..5669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     412..431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     665..682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  816..1040
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1107..1640)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1829..1834,1835..1851,1852..1857))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2253..2286)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  2364..2529
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        2914..3016
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3017..3676
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3083..3102)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3976..4430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4063..4082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4272..4293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4456..4560
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4561..4629,4630..5421)
                     /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(4779..4798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5592..5669,1..511)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
       61 aaatactatt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      541 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      601 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      661 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
      721 gcatctgtgc ggtatttcac accgcataga ccagccgcgt aacctggcaa aatcggttac
      781 ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa
      841 ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa
      901 actgaaatca gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc
      961 aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg
     1021 gcatggtaaa gactatattc gcggcgttgt gacaatttac cgaacaactc cgcggccggg
     1081 aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat atcaaagtgc
     1141 atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc gtcaccgtaa
     1201 tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt gaggagattg
     1261 atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag atcatagata
     1321 tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca
     1381 accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag caagttcccg
     1441 aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa ctcacgaccg
     1501 aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct acatgtgcga
     1561 atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg ctgcgtaaca
     1621 tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     1681 acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc
     1741 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     1801 gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg
     1861 ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga
     1921 agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg
     1981 tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctt
     2041 gcttcaggag atcggtagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga
     2101 tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc
     2161 tagctatagt tctagtggtt ggctacagct ttgtttgtac tatcaacagg ttgaactgct
     2221 gatcaacaga tcctctacgc ggccgcggta ccataacttc gtatagcata cattatacga
     2281 agttatctgg cctaggttaa ttaaacgctc tatggtctaa agtttaaact ggatactatt
     2341 gcacgtttca atcaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt
     2401 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     2461 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     2521 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc
     2581 catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa aatttttctg
     2641 tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt tgcagcctga
     2701 actgaacagg agggacagct gatagaaaca gaagccactg gagcacctca aaaacaccat
     2761 catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg aataaatacc
     2821 tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg ataccgggaa
     2881 gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt
     2941 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     3001 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     3061 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     3121 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     3181 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat
     3241 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     3301 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     3361 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     3421 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     3481 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     3541 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     3601 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     3661 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggttgcta
     3721 cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa attcgacccg
     3781 gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg gtttaccggt
     3841 ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg ccagtttgct
     3901 caggctctcc ccgtggaggt aataattgac gatatgatca tttattctgc ctcccagagc
     3961 ctgatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4021 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4081 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4141 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4201 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4261 cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4321 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4381 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4441 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4501 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4561 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4621 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4681 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4741 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4801 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4861 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4921 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4981 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5041 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5101 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5161 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5221 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5281 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5341 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5401 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5461 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5521 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5581 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5641 ccaccgctac cagcggtggt ttgtttgcc
//
