LOCUS       Exported                5668 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pBIG2abcde - Step2 vector abcde (biGBac system for expression of 
            protein complexes in insect cells).
ACCESSION   .
VERSION     .
KEYWORDS    pBIG2abcde
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  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 5668)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5668
                     /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"
     CDS             406..1065
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(472..491)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1364..1818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1451..1470)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1660..1681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1844..1948
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1949..2809
                     /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(2167..2186)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2980..3568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3469..3488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3722..3739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3873..4097
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4164..4697)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(4886..4914)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    5310..5343
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     mobile_element  5421..5586
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
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 ctgtagcggc
     1381 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1441 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1501 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1561 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1621 gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg
     1681 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1741 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1801 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1861 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1921 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1981 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2041 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2101 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2161 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2221 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2281 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2341 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2401 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2461 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2521 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2581 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2641 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2701 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2761 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2821 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2881 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3661 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3721 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3781 tctgtgcggt atttcacacc gcatagacca gccgcgtaac ctggcaaaat cggttacggt
     3841 tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg
     3901 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
     3961 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
     4021 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca
     4081 tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     4141 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     4201 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     4261 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     4321 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag
     4381 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
     4441 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     4501 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
     4561 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
     4621 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     4681 ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     4741 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat
     4801 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     4861 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt
     4921 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca gcagcgaagt
     4981 cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca
     5041 ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc tgcccttgct
     5101 tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga ccccggatga
     5161 agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc aggactctag
     5221 ctatagttct agtggttggc tacagctttg tttgtactat caacaggttg aactgctgat
     5281 caacagatcc tctacgcggc cgcggtacca taacttcgta tagcatacat tatacgaagt
     5341 tatctggcct aggttaatta aacgctctat ggtctaaagt ttaaacactg acattgactt
     5401 ggtttcaatc aggagatccg aaccagataa gtgaaatcta gttccaaact attttgtcat
     5461 ttttaatttt cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt
     5521 ccctaaataa tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg
     5581 cccacagcgg ggcatttttc ttcctgttat gtttttaatc aaacatcctg ccaactccat
     5641 gtgacaaacc gtcatcttcg gctacttt
//
