LOCUS       sequence-390120-        8789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mini-Agrin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8789)
  TITLE     Jeff Miner Lab Plasmids
REFERENCE   2  (bases 1 to 8789)
  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"
COMMENT     Sequence Label: pCI_E01_mini-mAgrin-Myc-His-WPRE__LG1
FEATURES             Location/Qualifiers
     source          1..8789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    42..325
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        516..895
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        896..1099
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1050..1070
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1235..1367
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1350..1369
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1411..1430
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1411..1429
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1530..1539
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3942..3962
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3987..4016
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4026..4055
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4065..4094
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4104..4133
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4143..4172
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4200..4217
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4254..4842
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4307..4327)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4725..4736)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4930..4948)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4985..5106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4996..5015)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5050..5069
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5351..5708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5351..5542
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5374..5394)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5559..5694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5739..6776
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             6755..6763
                     /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    6800..6848
                     /note="synthetic polyadenylation signal"
     promoter        7004..7108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7109..7177,7178..7969)
                     /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(7327..7346)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(8127..8715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8207..8226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca
       61 cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta
      121 agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta
      181 tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc
      241 gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact
      301 tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg
      361 cgtagaggat cgagatcttc aatattggcc attagccata ttattcattg gttatatagc
      421 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
      481 ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta
      541 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      601 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      661 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      721 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
      781 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      841 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      901 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
      961 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1021 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     1081 ggtctatata agcagagctg gtttagtgaa ccgtcagatc actagaagct ttattgcggt
     1141 agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt
     1201 aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     1261 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1321 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     1381 gttcaattac agctcttaag gctagagtat taatacgact cactataggg ctagcaaagc
     1441 gatcgccgaa ctcacctatc tccccaacac ctaataacat tcaatcactc tttccactaa
     1501 ccacctatct acatcaccac ccaccagccg ccaccatggt ccgcccgcgg ctgagcttcc
     1561 cggccccgct gctgccgctg ctgctgctcc tggcggcggc cgcgccagcc gtgccgggcg
     1621 ccagcgggac gtgcccggag cgcgccttgg agcggcgcga ggaggaggcg aacgtggtgc
     1681 tcaccggcac agtggaggag atcctcaacg tggacccggt gcagcacacc tactcctgca
     1741 aggttcgagt gtggcggtac ttgaaaggca aagatgtggt ggcccaagag agcctgctgg
     1801 atggtggcaa caaggtggta attggtggct ttggagaccc cctcatctgc gacaaccagg
     1861 tgtccaccgg ggacactaga atcttctttg tgaacccagc accaccctac ctgtggccag
     1921 cccacaagaa tgagttgatg ctcaactcca gccttatgcg gatcactttg cggaacctgg
     1981 aagaggtgga gttctgtgta gaagacaaac ctgggatcca cttcacagca gcgccttcta
     2041 tgcctcctga tgtgtgcagg ggaatgttat gtggctttgg tgctgtgtgt gaacctagtg
     2101 ttgaggatcc aggccgggcc tcctgtgtgt gcaagaagaa tgtctgccct gctatggtag
     2161 ctcctgtgtg tggctcagat gcttccacct atagcaacga gtgtgagcta cagcgtgcac
     2221 agtgcaacca gcaacggcgc atccgcctac tccgccaagg gccatgtggg tcccgggacc
     2281 cctgtgccaa tgtgaccgct agcactccag gcccgcaacg gcccccaaag tcctgtgatt
     2341 cccagccttg cctccacgga ggtacctgcc aggacctgga ttctggcaag ggtttcagct
     2401 gcagctgtac tgcaggcagg gctggcactg tctgtgagaa agtgcagctc ggagaccatc
     2461 cctgctcacc taacccctgc catggcgggg ccctctgcca ggccctggag gctggcgtgt
     2521 tcctctgtca gtgtccacct ggccgctttg gcccaacttg tgcagatgaa aagaacccct
     2581 gccaaccgaa cccctgccac gggtcagccc cctgccatgt gctttccagg ggtggggcca
     2641 agtgtgtgtg ccccctggga cgcagtggtt ccttctgtga gacagtcctg gagaatgctg
     2701 gctcccggcc cttcctggct gactttaatg gcttctccta cctggaactg aaaggcttgc
     2761 acaccttcga gagagaccta ggggagaaga tggcgctgga gatggtgttc ttggctcgtg
     2821 ggcccagtgg cttactcctc tacaatgggc agaagacgga tggcaagggg gactttgtat
     2881 ccctggccct gcataaccgg cacctagagt tccgctatga ccttggcaag ggggctgcaa
     2941 tcatcaggag caaagagccc atagccctgg gcacctgggt tagggtattc ctggaacgaa
     3001 atggccgcaa gggtgccctt caagtgggtg atgggccccg tgtgctaggg gaatctccgg
     3061 tcccgcacac catgctcaac ctcaaggagc ccctctatgt ggggggagct cctgacttca
     3121 gcaagctggc tcggggcgct gcagtggcct ccggctttga tggtgccatc cagctggtgt
     3181 ctctaagagg ccatcagctg ctgactcagg agcatgtgtt gcgggcagta gatgtagcgc
     3241 cttttgcagg ccacccttgt acccaggccg tggacaaccc ctgccttaat gggggctcct
     3301 gtatcccgag ggaagccact tatgagtgcc tgtgtcctgg gggcttctct gggctgcact
     3361 gcgagaaggg gatagttgag aagtcagtgg gggacctaga aacactggcc tttgatgggc
     3421 ggacctacat cgagtacctc aatgctgtga ctgagagtga gaaagcgctg cagagcaacc
     3481 actttgagct gagcttacgc actgaggcca cgcaggggct ggtgctgtgg attggaaagg
     3541 ttggagaacg tgcagactac atggctctgg ccattgtgga tgggcaccta caactgagct
     3601 atgacctagg ctcccagcca gttgtgctgc gctccactgt gaaggtcaac accaaccgct
     3661 ggcttcgagt cagggctcac agggagcaca gggaaggttc ccttcaggtg ggcaatgaag
     3721 cccctgtgac tggctcttcc ccgctgggtg ccacacaatt ggacacagat ggagccctgt
     3781 ggcttggagg cctacagaag cttcctgtgg ggcaggctct ccccaaggcc tatggcacgg
     3841 gttttgtggg ctgtctgcgg gacgtggtag tgggccatcg ccagctgcat ctgctggagg
     3901 acgctgtcac caaaccagag ctaagaccct gccccactct cgaggatctg tactttcaga
     3961 gcatcgatgg aagtggtaaa gctatggagc aaaagctcat ttctgaagag gacttgaatg
     4021 aaatggagca aaagctcatt tctgaagagg acttgaatga aatggagcaa aagctcattt
     4081 ctgaagagga cttgaatgaa atggagcaaa agctcatttc tgaagaggac ttgaatgaaa
     4141 tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagagcttg ggcgacctcc
     4201 atcatcacca tcaccactag gggatatcag ctgaattcta gaaattcatc gacaatcaac
     4261 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     4321 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     4381 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     4441 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     4501 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     4561 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     4621 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     4681 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4741 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4801 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcccaggttc cttgggccgc
     4861 gactctaggg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     4921 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     4981 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     5041 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     5101 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     5161 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt
     5221 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5281 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     5341 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     5401 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5461 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5521 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5581 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5641 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5701 tttgcaaaaa gctcgattct tctgacacta gcgccaccat gaagaagccc gaactcaccg
     5761 ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac ctgatgcagt
     5821 tgtcggaggg cgaagagagc cgagccttca gcttcgatgt cggcggacgc ggctatgtac
     5881 tgcgggtgaa tagctgcgct gatggcttct acaaagaccg ctacgtgtac cgccacttcg
     5941 ccagcgctgc actacccatc cccgaagtgt tggacatcgg cgagttcagc gagagcctga
     6001 catactgcat cagtagacgc gcccaaggcg ttactctcca agacctcccc gaaacagagc
     6061 tgcctgctgt gttacagcct gtcgccgaag ctatggatgc tattgccgcc gccgacctca
     6121 gtcaaaccag cggcttcggc ccattcgggc cccaaggcat cggccagtac acaacctggc
     6181 gggatttcat ttgcgccatt gctgatcccc atgtctacca ctggcagacc gtgatggacg
     6241 acaccgtgtc cgccagcgta gctcaagccc tggacgaact gatgctgtgg gccgaagact
     6301 gtcccgaggt gcgccacctc gtccatgccg acttcggcag caacaacgtc ctgaccgaca
     6361 acggccgcat caccgccgta atcgactggt ccgaagctat gttcggggac agtcagtacg
     6421 aggtggccaa catcttcttc tggcggccct ggctggcttg catggagcag cagactcgct
     6481 acttcgagcg ccggcatccc gagctggccg gcagccctcg tctgcgagcc tacatgctgc
     6541 gcatcggcct ggatcagctc taccagagcc tcgtggacgg caacttcgac gatgctgcct
     6601 gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc ggtcgcacac
     6661 aaatcgctcg ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg ctggccgaca
     6721 gcggcaaccg ccggcccagt acacgaccgc gcgctaagga ggtaggtcga gtttagactc
     6781 tagccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     6841 tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc ttgtccagat
     6901 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     6961 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7021 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7081 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7141 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7201 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7261 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttcaa
     7321 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7381 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7441 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7501 tgcggccaac ttacttctga caactatcgg aggaccgaag gagctaaccg cttttttgca
     7561 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7621 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7681 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7741 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7801 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7861 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7921 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat tcgaaatgac
     7981 cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     8041 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     8101 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     8161 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     8221 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     8281 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8341 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8401 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8461 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8521 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     8581 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8641 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8701 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8761 gaaaactcac gttaagggat tttggtcat
//
