LOCUS       sequence-177323-        6263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of mouse CSPG5 short isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6263)
  TITLE     Rathjen lab plasmids
REFERENCE   2  (bases 1 to 6263)
  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: pFlagCMV-mCALEBshort
FEATURES             Location/Qualifiers
     source          1..6263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(101..120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     311..332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     595..617
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     610..626
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        787..1166
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1167..1370
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1321..1341
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1401..1445
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1446..1469
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3073..3695
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3272..3291)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3719..3739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3724..4053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3904..4039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4091..4110)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4092..4110)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4124..4140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4124..4140)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4137..4159)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4148..4164
                     /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)."
     promoter        complement(join(4172..4178,4179..4196,4197..4202))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4217..4238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4355..4372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4526..5114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4606..4625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5285..6076,6077..6145))
                     /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     5908..5927
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6146..6250)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
       61 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      421 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat
      481 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      541 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      601 cacgacgttg taaaacgacg gccagtgcca agctgatcta tacattgaat caatattggc
      661 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
      721 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
      781 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      841 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      901 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      961 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     1021 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
     1081 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
     1141 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
     1201 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     1261 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
     1321 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     1381 accgtcagaa ttaattcacc atgtctgcac ttctgatcct agctcttgtt ggagctgcag
     1441 ttgctgacta caaagacgat gacgacaagc ttgcggccgc gaattcatcg atagatctga
     1501 tatcggtacc agtcgactct agaggatccg taccggcacg ggaaacgggc agtgcgatcg
     1561 aggctgaaga gctggtgagg agcagcctgg catgggagtc acgtgccaat gacacgcggg
     1621 aggaagccgg cctgccagca gctggggaag atgagacctc gtggacagag cggggcagtg
     1681 agatggctgc ggtgggccct ggggtcgggc cagaggaggc actagaggca tcggctgcag
     1741 tgactggcac tgcctggcta gaggcagatg gcccaggcct gggtggagtg actgcagagg
     1801 ctggcagtgg cgacgcccag acccttccag ctacgctcca ggctcctgat gaggcccttg
     1861 ggtcatctac aatgccccct gccatccctg aggctactga aaccagtgga cctccctccc
     1921 ctgctgtcca tgataagcct agtgtaggcc ctgaactccc taaagagatc cccttggagg
     1981 ttcggctgaa cctgggaggc agcacaccag agcccacttt tccccttcag ggcactctcg
     2041 agacccaacc agcctcagat ataattgaca ttgattactt tgaaggattg gatagtgagg
     2101 gtcgtggtgc agacatgggc agcttcccgg ggtcaccagg aacctcagaa aatcaccctg
     2161 ataccgaagg agagacccct tcctggagcc tgcttgattt gtatgatgac ttcacccctt
     2221 ttgatgagtc tgatttctac cccaccacat ccttctatga tgatttggaa gaggaggaag
     2281 aagaggagga ggataaggat acagtaggag gtggagacct ggaagatgaa aacgaccttc
     2341 tcctgccctc tcaaaagcct ggtgtggggc ctgggacagg acagcccaac aaccggtggc
     2401 atgctgttcc cccacagcat actctgggga tggtacctgg cagcagcatc tctcttaggc
     2461 cccgccccgg agatccaggc aaggacctgg cctcaggaga aaatggcaca gagtgccgag
     2521 ttggcttcgt caggcacaat ggctcctgcc ggtcagtctg tgacctcttt ccgagttact
     2581 gtcacaacgg cggccagtgc tacctggtgg agaacatagg ggctttctgc aggtgtaaca
     2641 cccaggacta catctggcac aaggggatgc gctgtgagtc catcatcacg gacttccagg
     2701 tgatgtgcgt ggccgttggc tcggctgctc tcgtgcttct cctcctgttc atgatgactg
     2761 tgttctttgc caagaagctc tatctgctca agactgagaa taccaagctg cggaggacca
     2821 ataaattccg gaccccatct gagctccaca acgacaactt ctccctctcc accattgccg
     2881 agggctctca tccaaatgac gaccccagcg ctccccacaa aatccaggac cctctcaagt
     2941 cccgcctgaa ggaggaagag tcctttaaca tccagaactc catgtcaccc aaacttgagg
     3001 gtggcaaagg tgaccaggat gacttggggg tgaactgtct gcagaataac ctaacctgag
     3061 actgaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3121 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3181 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3241 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3301 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     3361 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     3421 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     3481 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     3541 cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg
     3601 ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca
     3661 tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg
     3721 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3781 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3841 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3901 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3961 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4021 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag
     4081 aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg
     4141 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4201 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4261 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4321 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4381 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4441 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4501 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4561 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4621 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4681 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4741 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4801 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4861 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4921 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4981 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5041 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5101 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5161 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5221 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5281 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5341 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5401 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5461 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5521 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5581 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5641 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5701 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5761 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5821 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5881 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5941 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6001 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6061 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6121 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6181 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6241 gggttccgcg cacatttccc cga
//
