LOCUS       Exported                4113 bp ds-DNA     circular SYN 12-OCT-2021
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 4113)
  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 4113)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 12, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: ""Journal Article""; journalName: ""Proc 
            Natl Acad Sci U S A. 2016 May 10""; volume: ""113(19)""; pages: 
            ""E2564-9. doi: 10.1073/pnas.1604935113. Epub 2016 Apr 25""
FEATURES             Location/Qualifiers
     source          1..4113
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        303..405
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             406..1065
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /note="color: #ccffcc"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(472..491)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
                     /note="color: black; sequence: GCAACTGACTGAAATGCCTC"
     primer_bind     complement(1354..1371)
                     /label=MP1.1
                     /note="color: black; sequence: gggcgcgtcccattcgcc; added: 
                     2020-10-09"
     rep_origin      1364..1371
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     1372..1396
                     /label=MP1.2
                     /note="color: black; sequence: gagttttcgttccactgagcgtcag;
                     added: 2020-10-09"
     rep_origin      1425..2013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     1914..1933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     2167..2184
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     mobile_element  2318..2542
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
                     /note="color: #ffe4c4"
     CDS             complement(2609..3142)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /note="color: #ccffcc"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3331..3359)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
                     /note="This reverse directional feature has 3 segments:
                     1: 3331 .. 3336 / #ffffff / -10
                     2: 3337 .. 3353 / #ffffff
                     3: 3354 .. 3359 / #ffffff / -35"
     primer_bind     3730..3754
                     /label=2.16
                     /note="color: black; sequence: agatcctctacgcggccgcggtacc;
                     added: 2021-10-12"
     primer_bind     complement(3737..3754)
                     /label=2.14
                     /note="color: black; sequence: ggtaccgcggccgcgtag; added: 
                     2021-10-12"
     protein_bind    3755..3788
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    3805..3827
                     /label=bigbac_A
                     /note="color: #00ccff"
     misc_feature    3828..3850
                     /label=bigbac_C
                     /note="color: #cc99ff"
     mobile_element  3866..4031
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
                     /note="color: #ffe4c4"
     primer_bind     4010..4032
                     /label=2.15
                     /note="color: black; sequence: ccaactattttgtccgcccacag;
                     added: 2021-10-12"
     primer_bind     complement(4010..4032)
                     /label=2.17
                     /note="color: black; sequence: ctgtgggcggacaaaatagttgg;
                     added: 2021-10-12"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa ctgaacagga gggacagctg atagaaacag
      121 aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca
      181 cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc agaataaata
      241 aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac
      301 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
      361 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
      421 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
      481 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
      541 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
      601 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
      661 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
      721 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
      781 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
      841 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
      901 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
      961 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
     1021 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     1081 tattggtgcc cttaaacgcc tggtgctacg cctgaataag tgataataag cggatgaatg
     1141 gcagaaattc gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc
     1201 cgcttatgtc tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg
     1261 cttctcaaat gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga
     1321 tatgatcatt tattctgcct cccagagcct gatggcgaat gggacgcgcc cgagttttcg
     1381 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1441 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1501 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1561 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1621 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1681 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1741 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1801 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1861 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1921 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1981 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2041 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2101 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2161 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     2221 acgcatctgt gcggtatttc acaccgcata gaccagccgc gtaacctggc aaaatcggtt
     2281 acggttgagt aataaatgga tgccctgcgt aagcgggtgt gggcggacaa taaagtctta
     2341 aactgaacaa aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt
     2401 aaactgaaat cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa
     2461 gcaaactctt cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa
     2521 gggcatggta aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg
     2581 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     2641 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     2701 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     2761 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     2821 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     2881 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     2941 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     3001 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     3061 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     3121 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     3181 taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa
     3241 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     3301 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac
     3361 tgggtttgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc
     3421 gaagtcgagg catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat
     3481 cgtcaggcat tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc
     3541 ttgcttcagg agatcggtag acctcggccg tcgcggcgct tgccggtggt gctgaccccg
     3601 gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac
     3661 tctagctata gttctagtgg ttggctacag ctttgtttgt actatcaaca ggttgaactg
     3721 ctgatcaaca gatcctctac gcggccgcgg taccataact tcgtatagca tacattatac
     3781 gaagttatct ggcctaggtt aattaaacgc tctatggtct aaagtttaaa ctggatacta
     3841 ttgcacgttt caatcaggag atccgaacca gataagtgaa atctagttcc aaactatttt
     3901 gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca
     3961 ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt
     4021 gtccgcccac agcggggcat ttttcttcct gttatgtttt taatcaaaca tcctgccaac
     4081 tccatgtgac aaaccgtcat cttcggctac ttt
//
