LOCUS       sequence-223837-        6567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the extracellular domain of CBLN3 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 6567)
  TITLE     Comoletti Lab Plasmids
REFERENCE   2  (bases 1 to 6567)
  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-CBLN3-Fc
FEATURES             Location/Qualifiers
     source          1..6567
                     /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             1601..1624
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2933..3555
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3132..3151)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3579..3599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3584..3913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3764..3899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3951..3970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3952..3970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3984..4000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3984..4000)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3997..4019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4008..4024
                     /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(4032..4038,4039..4056,4057..4062))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4077..4098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4215..4232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4386..4974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4466..4485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5145..5936,5937..6005))
                     /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     5768..5787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6006..6110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6137..6565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6224..6243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6434..6455
                     /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 agagggcagc gagcctgtgc tgctggaggg agagtgtctg gtggtgtgcg agcctggcag
     1141 agctgctgcc ggaggacctg gaggagccgc tctgggcgaa gcccctcctg gaagagtggc
     1201 cttcgccgcc gtgagaagcc accaccacga acctgccgga gagacaggca acggcaccag
     1261 cggcgccatc tacttcgacc aggtgctggt gaacgagggc ggcggcttcg acagagccag
     1321 cggcagcttc gtggcccccg tgagaggcgt gtacagcttc agattccacg tggtgaaggt
     1381 gtacaacaga cagaccgtcc aggtgagcct gatgctgaac acctggcccg tgatcagcgc
     1441 cttcgccaac gaccccgacg tgaccagaga ggccgccacc agcagcgtgc tgctgcctct
     1501 ggaccccggc gacagagtga gcctgagact gagaagaggc aacctgctgg gcggctggaa
     1561 gtacagcagc ttcagcggct tcctgatctt ccccctgggt ctagaggtgc tcttccaggg
     1621 gcctgatccc gatcccgagg gtgagtacta agcttcagcg ctcctgcctg gacgcatccc
     1681 ggctatgcag ccccagtcca gggcagcaag gcaggccccg tctgcctctt cacccggagg
     1741 cctctgcccg ccccactcat gctcagggag agggtcttct ggctttttcc ccaggctctg
     1801 ggcaggcaca ggctaggtgc ccctaaccca ggccctgcac acaaaggggc aggtgctggg
     1861 ctcagacctg ccaagagcca tatccgggag gaccctgccc ctgacctaag cccaccccaa
     1921 aggccaaact ctccactccc tcagctcgga caccttctct cctcccagat tccagtaact
     1981 cccaatcttc tctctgcaga gcccaaatct tgtgacaaaa ctcacacatg cccaccgtgc
     2041 ccaggtaagc cagcccaggc ctcgccctcc agctcaaggc gggacaggtg ccctagagta
     2101 gcctgcatcc agggacaggc cccagccggg tgctgacacg tccacctcca tctcttcctc
     2161 agcacctgaa ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac
     2221 cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga
     2281 ccctgaggtc aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa
     2341 gccgcgggag gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca
     2401 ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc
     2461 ccccatcgag aaaaccatct ccaaagccaa aggtgggacc cgtggggtgc gagggccaca
     2521 tggacagagg ccggctcggc ccaccctctg ccctgagagt gaccgctgta ccaacctctg
     2581 tccctacagg gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggatgagc
     2641 tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg
     2701 ccgtggagtg ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc
     2761 tggactccga cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc
     2821 agcaggggaa cgtcttctca tgctccgtga tgcatgaggc tctgcacaac cactacacgc
     2881 agaagagcct ctccctgtct ccgggtaaat gagtgcgacg gccgcgactc tagggtggca
     2941 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     3001 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     3061 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     3121 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     3181 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     3241 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     3301 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     3361 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa
     3421 ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc
     3481 ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta
     3541 gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg
     3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3661 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3721 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3781 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3841 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3901 aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt ccctatagtg
     3961 agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4021 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4081 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4141 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4201 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4261 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4321 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacatagtt cttgaagtgg
     4801 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5221 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5281 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5341 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5401 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5461 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5521 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5581 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5641 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5701 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5761 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5821 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5881 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5941 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6001 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6061 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6121 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6181 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6241 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6301 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6361 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6421 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6481 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6541 atttaacaaa aatttaacgc gaatttt
//
