LOCUS       sequence-92020-e        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for E. coli expression of Patronin CC truncation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  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 6764)
  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: pET-21a-GFP-Patronin CC 1288–1457
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..202
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             212..229
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(233..235,236..238,239..949)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(233..235,236..238,239..949)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(278..299)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(539..558)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     886..907
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    959..983
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1528..1560)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             1568..1590
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1605..1629
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(1629..1648)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1630..1648)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1732..1751)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1957..2034
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2035..3117
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2054..2073)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3926..4117
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4111..4133)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4219..4361
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4274..4293
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4376..4393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4547..5135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4627..4646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5306..6097,6098..6166))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5929..5948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6167..6271)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6298..6753)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6435..6456)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6647..6666
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcatgggcag
      181 cagccatcat catcatcatc acagcagcgg cctggtgccg cgcggcagcc atatggtgag
      241 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      301 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      361 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      421 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
      481 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      541 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      601 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      661 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
      721 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
      781 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      841 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
      901 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc accggtatac
      961 aagtttgtac aaaaaagcag gctccgcggc cgctagcagc aacaacgcag aggcggccaa
     1021 gaacaaggcg ctggtcatcg gagcagattc caccaatttg gatccggaat ctgttgatga
     1081 gatggagcgg cgcaaggaga aaatcatgct gctgtctttg caacgtcgcc agcaacagga
     1141 ggaggcgaag gcgcgcaaag agattgaggc ttcccagaag cgagaaaagg agcgcgaaaa
     1201 ggaggaggaa cgagcgcgga aaaaggagga gcaaatggca cggcgagcgg ccattttgga
     1261 gcagcacaga ctcaagaaag ccattgaaga ggccgagcga gagggtaaaa ccctggatcg
     1321 gcccgatctg cacgtgaagc tgcaatcgca ttcatccacc tcaacgaccc cacgactgag
     1381 gcagcagcgc accacgcgtc ccaggcctaa gacaattcac gtggacgatg ccagcgtgga
     1441 catcagcgag gcttcaagca tctctagtcg gggcaaaaaa ggctcaagct cgaatccggc
     1501 tgctaacaaa gcccgaaagg atccgcgacc catttgctgt ccaccagtca tgctagccat
     1561 atgtatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct
     1621 cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg
     1681 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     1741 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     1801 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     1861 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     1921 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
     1981 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     2041 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     2101 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     2161 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     2221 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     2281 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     2341 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     2401 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     2461 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     2521 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     2581 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     2641 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     2701 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     2761 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     2821 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     2881 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     2941 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     3001 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3061 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3121 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     3181 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     3241 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     3301 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     3361 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     3421 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
     3481 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     3541 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     3601 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     3661 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     3721 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     3781 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     3841 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     3901 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     3961 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     4021 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     4081 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4141 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4201 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     4261 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg
     4321 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     4381 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4441 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4501 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4561 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4621 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4681 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4741 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4801 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4861 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4921 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4981 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5041 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5101 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5161 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5221 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5281 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5341 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5401 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5461 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5521 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5581 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     5641 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5701 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5761 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5821 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5881 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5941 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6001 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6061 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6121 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6181 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6241 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgaaa
     6301 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6361 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     6421 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6481 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6541 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6601 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6661 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6721 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gcca
//
