LOCUS       sequence-457380-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBIG2abcde - Step2 vector abcde (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: pBIG2abcde
FEATURES             Location/Qualifiers
     source          1..5669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        473..575
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             576..1235
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(642..661)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1535..1989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1622..1641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1831..1852
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2015..2119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2120..2188,2189..2980)
                     /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(2338..2357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3151..3739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3640..3659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3893..3910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4044..4268
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4335..4868)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5057..5062,5063..5079,5080..5085))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(5481..5514)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  join(5592..5669,1..88)
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc
       61 cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa
      121 tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca
      181 cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca
      241 acgcttattt gcagcctgaa ctgaacagga gggacagctg atagaaacag aagccactgg
      301 agcacctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca cccgacgcac
      361 tttgcgccga ataaatacct gtgacggaag atcacttcgc agaataaata aatcctggtg
      421 tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac gttgatcggc
      481 acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg
      541 agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
      601 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
      661 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
      721 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
      781 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
      841 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
      901 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
      961 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     1021 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     1081 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     1141 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     1201 aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     1261 cttaaacgcc tggttgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt
     1321 cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt cgttaaatag ccgcttatgt
     1381 ctattgctgg tttaccggtt tattgactac cggaagcagt gtgaccgtgt gcttctcaaa
     1441 tgcctgaggc cagtttgctc aggctctccc cgtggaggta ataattgacg atatgatcat
     1501 ttattctgcc tcccagagcc tgatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1561 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1621 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1681 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1741 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1801 cctttgacgt tggagtccac gttcttaata gtggactctt gttccaaact ggaacaacac
     1861 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1921 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     1981 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2041 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2101 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2161 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2221 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2281 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2341 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2401 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2461 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2521 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2581 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2641 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2701 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2761 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2821 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     2881 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2941 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3001 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3061 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactattct
     3301 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3781 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3841 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3901 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3961 tatttcacac cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata
     4021 aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact gaacaaaata
     4081 gatctaaact atgacaataa agtcttaaac tagacagaat agttgtaaac tgaaatcagt
     4141 ccagttatgc tgtgaaaaag catactggac ttttgttatg gctaaagcaa actcttcatt
     4201 ttctgaagtg caaattgccc gtcgtattaa agaggggcgt ggccaagggc atggtaaaga
     4261 ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg
     4321 gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc
     4381 cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg
     4441 tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg
     4501 gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta
     4561 cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg
     4621 tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag
     4681 tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga
     4741 gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata
     4801 cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctg
     4861 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     4921 cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc
     4981 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
     5041 acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg tttgtgcctt
     5101 catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag tcgaggcatt
     5161 tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc
     5221 ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgcccttgc ttcaggagat
     5281 cggtagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg aagtggttcg
     5341 catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta gctatagttc
     5401 tagtggttgg ctacagcttt gtttgtacta tcaacaggtt gaactgctga tcaacagatc
     5461 ctctacgcgg ccgcggtacc ataacttcgt atagcataca ttatacgaag ttatctggcc
     5521 taggttaatt aaacgctcta tggtctaaag tttaaacact gacattgact tggtttcaat
     5581 caggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt
     5641 tcgtattagc ttacgacgct acacccagt
//
