LOCUS       sequence-123551-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 400 amino acids of mouse BICD2 codon optimized for Sf9 
            expression with a N-terminal GFP tag and a TEV-cleavable His-ZZ tag 
            for purification (GFP-BICD2N).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  AUTHORS   Schlager MA, Hoang HT, Urnavicius L, Bullock SL, Carter AP
  TITLE     In vitro reconstitution of a highly processive recombinant human 
            dynein complex.
  JOURNAL   EMBO J. 2014 Sep 1;33(17):1855-68. doi: 10.15252/embj.201488792. 
            Epub 2014 Jul 1.
  PUBMED    24986880
REFERENCE   2  (bases 1 to 6960)
  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: 
            "10.15252/embj.201488792"; journalName: "EMBO J"; date: "2014-09-1- 
            1"; volume: "33"; issue: "17"; pages: "1855-68"
COMMENT     Sequence Label: GFP-BICD2N
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        74..165
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     90..107
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             232..255
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             298..471
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAYYEILHLPNLNEGQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             472..645
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAEKLNDAQAPK"
     CDS             703..723
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             745..765
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1947..1958
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2795..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2832..2851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2886..2905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3050..3215
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     3222..3927
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3310..3682
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3339..3927)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     rep_origin      4117..4705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4606..4625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_recomb     4957..5784
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4957..5081
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5119..5724)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     mobile_element  5813..6036
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6103..6636)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6825..6830,6831..6847,6848..6853))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(6924..6957)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 taactataac ggtcctaagg tagcgagttt aaacactagt atcgattcgc gacctactcc
       61 ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta
      121 ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca
      181 taccgtccca ccatcgggcg cggatctcgg tccgaaacca tgtcgtacta ccatcaccat
      241 caccatcacc atcacggttc tatgtgcatg gcaggccttg cgcaacacga tgaagccgtg
      301 gacaacaaat tcaacaaaga acaacaaaac gcgtactatg agatcttaca tttacctaac
      361 ttaaacgaag gacaacgaaa cgccttcatc caaagtttaa aagatgaccc aagccaaagc
      421 gctaaccttt tagcagaagc taaaaagcta aatgatgctc aggcgccgaa agtagacaac
      481 aaattcaaca aagaacaaca aaacgcgttc tatgagatct tacatttacc taacttaaac
      541 gaagaacaac gaaacgcctt catccaaagt ttaaaagatg acccaagcca aagcgctaac
      601 cttttagcag aagctgaaaa gctaaatgat gctcaggcgc cgaaagtaga cgcaaattcc
      661 gcggggaagt caaccgatta tgatattcca actactgcta gcgagaacct atatttccag
      721 ggagattatg atatccctac gactgagaat ctttattttc agggcatgtc taaaggtgaa
      781 gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac
      841 aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa
      901 tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttcggt
      961 tatggtgttc aatgttttgc gagataccca gatcatatga aacaacatga ctttttcaag
     1021 tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac
     1081 tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta
     1141 aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga atacaactat
     1201 aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc
     1261 aaaattagac acaacattga agacggttct gttcaattag ctgaccatta tcaacaaaat
     1321 actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct
     1381 gccttatcca aggatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact
     1441 gctgctggta ttaccggttc tatgtcagct ccatcagaag aagaggaata tgctaggttg
     1501 gtcatggaag cccagcccga gtggctcagg gctgaagtta agcgcctgtc tcacgagctc
     1561 gccgagacaa ccagagagaa gatccaagca gctgaatatg gcctcgctgt tttggaggag
     1621 aaacaccaac tgaaactcca gttcgaggag ctggaggttg actacgaggc cattaggtca
     1681 gaaatggagc aactgaaaga agccttcggc caagcacata ccaatcataa gaaggttgcc
     1741 gcagacggag agtccagaga agaatcactg atccaggagt ctgctagcaa ggagcagtat
     1801 tacgtgagga aagtcctgga gctgcagacc gagttgaagc aactcaggaa cgttctgacc
     1861 aacacccaat ctgaaaacga gagattgaca agcgtggccc aagagctcaa ggaaatcaat
     1921 cagaacgtcg agattcaacg cggtagattg agggacgaca tcaaagaata caaattcagg
     1981 gaggcaagac tcctccaaga ctactcagag ctggaagagg agaacatctc tctgcagaaa
     2041 caagtctcag tgctcaggca aaatcaggtg gagttcgaag gactgaagca cgaaatcaag
     2101 cgcctcgaag aggagacaga gtacctgaac tcccaactgg aggacgctat cagactgaag
     2161 gagatttcag aaagacagct cgaagaagcc ctggaaacac tgaagacaga aagggagcag
     2221 aagaataacc tccgcaagga gttgagccac tatatgtcta tcaacgattc cttctacact
     2281 tcacatctcc aggtcagctt ggacggactg aagttcagcg acgacactgt taccgccgag
     2341 cccaacaatg atgccgaagc tctcgtgaac ggattcgaac atagcggcct ggtcaaatcc
     2401 tcattggata acaagactag cactccacgc aaggatggtc tggctcctcc ctctccttcc
     2461 ttggtgtccg atctgttgag cgagttgcac atcagcgaaa tccagaagct caagcagcag
     2521 ctcgtccaaa tggagcgcga gaaggtgggt ctgctggcca cattgcaaga tactcagaaa
     2581 cagttggagc aggctagagg taccttgtca gagcagcatg agaaggtgaa caggttgacc
     2641 gagaatctga gcgctctgag gtaatgagct tgtcgagaag tactagagga tcataatcag
     2701 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2761 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2821 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2881 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgct
     2941 tgagcctaga agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     3001 ccgctgagca ataactatca taacccctag ggtataccca tctaattgga accagataag
     3061 tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac
     3121 acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca
     3181 cccctcccag ttcccaacta ttttgtccgc ccacaaccgg tggaggaaat tctccttgaa
     3241 gtttccctgg tgttcaaagt aaaggagttt gcaccagacg cacctctgtt cactggtccg
     3301 gcgtattaaa acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt
     3361 tgttgattta cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta
     3421 gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat
     3481 attaaatcct caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt
     3541 tccgaaccga tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa
     3601 tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt
     3661 tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac
     3721 aattgactcg acgtaaacac gttaaataga gcttggacat atttaacatc gggcgtgtta
     3781 gctttattag gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa
     3841 gacgattttg ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc
     3901 aaatttgtag ttgagctttt tggaattacc ggttgacttg ggtcaactgt cagaccaagt
     3961 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4021 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4081 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4141 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4201 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4261 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4321 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4381 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4441 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4501 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4561 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4621 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4681 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4741 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt gacttgggtc
     4801 gctcttcctg tggatgcgca ggtatgtaca ggaagaggtt tatactaaac tgttacattg
     4861 caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct ctgacgcatt
     4921 tctacaacca cgactctaag tgtgtgggtg aagtcatgca tcttttaatc aaatcccaag
     4981 atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt cgacaagctc tgtccgtttg
     5041 ctggcaactg caagggtctc aatcctattt gtaattattg aataataaaa caattataaa
     5101 tgtcaaattt gttttttatt aacgatacaa accaaacgca acaagaacat ttgtagtatt
     5161 atctataatt gaaaacgcgt agttataatc gctgaggtaa tatttaaaat cattttcaaa
     5221 tgattcacag ttaatttgcg acaatataat tttattttca cataaactag acgccttgtc
     5281 gtcttcttct tcgtattcct tctctttttc atttttctct tcataaaaat taacatagtt
     5341 attatcgtat ccatatatgt atctatcgta tagagtaaat tttttgttgt cataaatata
     5401 tatgtctttt ttaatggggt gtatagtacc gctgcgcata gtttttctgt aatttacaac
     5461 agtgctattt tctggtagtt cttcggagtg tgttgcttta attattaaat ttatataatc
     5521 aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg gcatagtacg cagcttcttc
     5581 tagttcaatt acaccatttt ttagcagcac cggattaaca taactttcca aaatgttgta
     5641 cgaaccgtta aacaaaaaca gttcacctcc cttttctata ctattgtctg cgagcagttg
     5701 tttgttgtta aaaataacag ccattgtaat gagacgcaca aactaatatc acaaactgga
     5761 aatgtctatc aatatatagt tgctgattgc gcagatgccc tgcgtaagcg ggtgtgggcg
     5821 gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact
     5881 agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact
     5941 tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa
     6001 gaggggcgtg gccaagggca tgtaaagact atattcgcgg cgttgtgaca atttaccgaa
     6061 caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg
     6121 gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc
     6181 gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc
     6241 atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac
     6301 tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc
     6361 cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact
     6421 acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc
     6481 tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc
     6541 gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact
     6601 gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac
     6661 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa
     6721 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     6781 aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac
     6841 aggcttatgt caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa
     6901 ccttgggcag cagcgaagtc gccataactt cgtatagcat acattatacg aagttatctg
//
