LOCUS       sequence-92001-e        7859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for baculovirus expression of CAMSAP2 CC+CKK in Sf9 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7859)
  AUTHORS   Hendershott MC, Vale RD
  TITLE     Regulation of microtubule minus-end dynamics by CAMSAPs and 
            Patronin.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Apr 22;111(16):5860-5. doi: 
            10.1073/pnas.1404133111. Epub 2014 Mar 26.
  PUBMED    24706919
REFERENCE   2  (bases 1 to 7859)
  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.1404133111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-04-22- 22"; volume: "111"; issue: "16"; pages: "5860-5"
COMMENT     Sequence Label: pFasBac+GFP-CAMSAP2 CC+CKK
FEATURES             Location/Qualifiers
     source          1..7859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     299..320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        484..588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(589..657,658..1449)
                     /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(807..826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1620..2208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2109..2128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2362..2379
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3904..3995
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3920..3937
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4101..4121
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4134..4136,4137..4847)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4134..4136,4137..4847)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4176..4197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4437..4456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4784..4805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5776..5784
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    7349..7483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7386..7405)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7440..7459
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7512..7677
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      421 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
      481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      781 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
      841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
      961 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1441 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1741 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1981 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2281 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2341 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2401 cggtattttc tccttacgca tctgtgcggt atttcacacc gcagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     4021 ccatcgggcg cggatctcgg tccgaaacca tgtcgtacta ccatcaccat caccatcacg
     4081 attacgatat cccaacgacc gaaaacctgt attttcaggg cgccatggat ccagtgagca
     4141 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     4201 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     4261 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4321 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     4381 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4441 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4501 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4561 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     4621 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     4681 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     4741 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     4801 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggag ctactgaatc
     4861 ataccgatgg aaaaagtagt ggaagcagtt ctcaaaaaac tacaccagaa ggctctgaac
     4921 ttaatattcc tcatgtggtt gcttgggcac aaattccaga agaaacaggg cttccacagg
     4981 gacgggacac tacccagctg ttggcctctg aaatggtgca tcttaggatg aaactagaag
     5041 aaaagaggcg tgctatagaa gcccagaaaa agaaaatgga agctgctttt accaaacaga
     5101 gacagaaaat gggaaggaca gcattcctta ctgtagtgaa aaagaaaggg gatgggatat
     5161 ctcctctacg agaggaagcg gcgggtgcag aagatgagaa agtatatact gatcgagcaa
     5221 aagaaaagga atcacaaaaa actgatggac aaaggagcaa gtcactggca gatataaaag
     5281 agagcatgga gaatcctcaa gccaaatggc taaagtctcc aactacacct attgatcctg
     5341 agaagcagtg gaacctggca agcccctcag aagaaacttt aaatgaagga gagattttag
     5401 aatataccaa atccattgaa aagttaaatt catccctgca ttttctacaa caagaaatgc
     5461 aacgcttgtc acttcagcag gagatgttaa tgcagatgag agagcaacaa tcttgggtga
     5521 tttcacctcc acaaccctct ccacagaaac agattcgaga ttttaagcct tctaagcagg
     5581 caggcctgtc atcagccatt gcaccattct cctcagactc ccctcgtcct actcacccat
     5641 ctccacagtc ttctaacagg aaaagtgcat ctttttctgt taaaagtcaa aggactccta
     5701 ggccaaatga gttaaaaata acacctttga atcgaacctt gacacctcct cggtctgtgg
     5761 atagccttcc tcggttaagg aggttttcac caagtcaagt tcctattcaa actaggtcat
     5821 ttgtatgttt tggggatgat ggagaacctc agttaaagga atccaaacct aaagaggaag
     5881 ttaaaaagga ggaattggaa tccaaaggga ctttggaaca gcgtggacat aatccagaag
     5941 aaaaggaaat caaacctttt gagtcaacag tctctgaagt cctatcactg cctgtcacag
     6001 agactgtatg tctgacacca aatgaggacc aattgaatca acccacagaa ccccctccta
     6061 aacccgtttt cccacccact gctccaaaaa atgttaatct gattgaagtt tccctctcag
     6121 atttgaaacc ccctgaaaag gctgatgtac ctgttgaaaa atatgatgga gaaagtgata
     6181 aagaacaatt tgatgatgac cagaaagtat gctgtggatt cttttttaag gatgatcaaa
     6241 aagcagaaaa tgatatggca atgaaacggg cagctttgtt ggagaaaaga ttaagaaggg
     6301 aaaaggaaac tcagctccgg aaacaacagt tggaagcaga aatggagcat aagaaggagg
     6361 aaacaaggcg taaaactgag gaagaacgtc agaagaaaga agatgagaga gcacgcagag
     6421 aatttattag gcaagaatat atgaggcgga aacaactgaa actaatggaa gatatggata
     6481 cagtaattaa accccgtcct caagtagtaa aacaaaaaaa acagcgacca aaatctattc
     6541 acagagatca tattgaatcc cccaaaacac caataaaggg tcctccagtc tctagccttt
     6601 ctttggcatc gctgaacacg ggtgataacg agagtgtaca ttcaggcaag aggacgccaa
     6661 gatcagagtc tgtagaaggc ttcttatctc caagtcgttg tggcagtcga aatggagaaa
     6721 aagactggga gaatgcatca acaacttctt cagtggcttc tggaacagaa tatacaggac
     6781 caaagctcta caaagaaccc agtgcaaaat ccaataagca cataatacaa aatgctttag
     6841 ctcattgctg tttggctgga aaagtaaatg aaggtcagaa gaaaaaaata ctggaggaaa
     6901 tggagaaatc agatgccaac aacttcttaa tcttgttccg ggattcagga tgccagttca
     6961 gatctttata cacttattgc ccagaaactg aagaaatcaa taaactgact gggataggcc
     7021 ctaaatctat cactaaaaaa atgattgaag gactttacaa atataattct gacaggaaac
     7081 agtttagcca catacccgct aaaactttat ctgccagtgt tgatgcaatt accattcata
     7141 gccatttatg gcagaccaaa agaccagtaa cacccaaaaa acttttaccc actaaggcat
     7201 agtcgaggca tgcggtacca agcttgtcga gaagtactag aggatcataa tcagccatac
     7261 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     7321 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     7381 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7441 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac tgcttgagcc
     7501 taggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt
     7561 tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata
     7621 atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc gcccacagcg
     7681 gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca tgtgacaaac
     7741 cgtcatcttc ggctactttt tctctgtcac agaatgaaaa tttttctgtc atctcttcgt
     7801 tattaatgtt tgtaattgac tgaatatcaa cgcttatttg cagcctgaat ggcgaatgg
//
