LOCUS       sequence-223842-        6561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the extracellular domain of CBLN1 with N-terminal FLAG
            tag and C-terminal hIgG tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6561)
  TITLE     Comoletti Lab Plasmids
REFERENCE   2  (bases 1 to 6561)
  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: pCMV6-XL4 FLAG-CBLN1-Fc
FEATURES             Location/Qualifiers
     source          1..6561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             956..1045
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             1046..1069
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1595..1618
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2927..3549
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3126..3145)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3573..3593)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3578..3907
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3758..3893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3945..3964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3946..3964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3978..3994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3978..3994)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3991..4013)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4002..4018
                     /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(4026..4032,4033..4050,4051..4056))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4071..4092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4209..4226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4380..4968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4460..4479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5139..5930,5931..5999))
                     /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     5762..5781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6000..6104)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6131..6559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6218..6237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6428..6449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 tttcaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatatcg ccaccatgga
      961 cagcaaaggt tcgtcgcaga aagggtcccg cctgctcctg ctgctggtgg tgtcaaatct
     1021 actcttgtgc cagggtgtgg tctccgacta caaagacgat gacgacaagg cggccgctca
     1081 aaacgagaca gagcccatcg tgctggaggg caagtgcctg gtggtgtgtg acagcaatcc
     1141 caccagcgac cctacaggca ccgccctggg catcagcgtg agaagcggca gcgccaaggt
     1201 ggccttcagc gccatcagaa gcaccaacca cgagcccagc gagatgagca acagaaccat
     1261 gatcatctac ttcgaccagg tgctggtgaa catcggcaac aacttcgaca gcgagagaag
     1321 caccttcatc gcccccagaa agggcatcta cagcttcaac ttccacgtgg tgaaggtgta
     1381 caacagacag accatccagg tgagcctgat gctgaacggc tggcctgtga tcagcgcctt
     1441 cgccggcgac caggacgtga ccagagaggc cgccagcaac ggcgtgctga tccagatgga
     1501 gaagggcgat agagcctacc tgaagctgga gagaggcaac ctgatgggcg gcaagtacag
     1561 caccttcagc ggcttcctgg tgttccccct gggtctagag gtgctcttcc aggggcctga
     1621 tcccgatccc gagggtgagt actaagcttc agcgctcctg cctggacgca tcccggctat
     1681 gcagccccag tccagggcag caaggcaggc cccgtctgcc tcttcacccg gaggcctctg
     1741 cccgccccac tcatgctcag ggagagggtc ttctggcttt ttccccaggc tctgggcagg
     1801 cacaggctag gtgcccctaa cccaggccct gcacacaaag gggcaggtgc tgggctcaga
     1861 cctgccaaga gccatatccg ggaggaccct gcccctgacc taagcccacc ccaaaggcca
     1921 aactctccac tccctcagct cggacacctt ctctcctccc agattccagt aactcccaat
     1981 cttctctctg cagagcccaa atcttgtgac aaaactcaca catgcccacc gtgcccaggt
     2041 aagccagccc aggcctcgcc ctccagctca aggcgggaca ggtgccctag agtagcctgc
     2101 atccagggac aggccccagc cgggtgctga cacgtccacc tccatctctt cctcagcacc
     2161 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     2221 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     2281 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     2341 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     2401 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     2461 cgagaaaacc atctccaaag ccaaaggtgg gacccgtggg gtgcgagggc cacatggaca
     2521 gaggccggct cggcccaccc tctgccctga gagtgaccgc tgtaccaacc tctgtcccta
     2581 cagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggat gagctgacca
     2641 agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg
     2701 agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact
     2761 ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg
     2821 ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac acgcagaaga
     2881 gcctctccct gtctccgggt aaatgagtgc gacggccgcg actctagggt ggcatccctg
     2941 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3001 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3061 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3121 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3181 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3241 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3301 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3361 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3421 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3481 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3541 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3601 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3661 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3721 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3781 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3841 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3901 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt
     3961 attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4021 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4081 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4141 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4201 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4261 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4321 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4381 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4441 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4501 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4561 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4621 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4681 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4741 aacaggatta gcagagcgag gtatgtaggc ggtgctacat agttcttgaa gtggtggcct
     4801 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4861 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4921 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4981 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5041 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5101 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5161 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5221 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5281 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5341 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5401 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5461 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5521 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5581 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5641 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5701 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5761 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5821 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5881 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5941 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6001 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6061 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6121 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6181 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6241 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6301 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6361 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6421 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6481 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6541 caaaaattta acgcgaattt t
//
