LOCUS       sequence-415705-        7891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to COMP5AP-AviTag-9xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7891)
  AUTHORS   Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL, 
            Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser 
            SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC
  TITLE     A Human IgSF Cell-Surface Interactome Reveals a Complex Network of 
            Protein-Protein Interactions.
  JOURNAL   Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 
            10.1016/j.cell.2020.07.025.
  PUBMED    32822567
REFERENCE   2  (bases 1 to 7891)
  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: 
            "10.1016/j.cell.2020.07"; journalName: "Cell"; date: "2020-08-20- 
            20"; volume: "182"; issue: "4"; pages: "1027-1043.e17"
COMMENT     Sequence Label: pD649-HAsp-HAVCR1-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1802..1837
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             1847..1870
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3530..3574
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3575..3601
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(3741..3758)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3747..3858
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3976..3996)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4020..4230
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4042..4238
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4089..4224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4151..4170
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4286..4333
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4352..4951
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4352..4371)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4848..4868
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5081..5202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5118..5137)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5172..5191
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5255..6046,6047..6115))
                     /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     5878..5897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6116..6207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6249..6268
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6274..6293
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6379..6967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6868..6887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7121..7138
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7236..7253
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7237..7253
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7709..7725
                     /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)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg cctctgtgaa agttggcgga gaagctggac ctagcgtgac
     1021 cctgccttgt cattattctg gcgccgtgac cagcatgtgc tggaatagag gcagctgcag
     1081 cctgttcacc tgtcagaatg gcatcgtgtg gaccaacggc acccacgtga cctacagaaa
     1141 ggacaccaga tacaagctgc tgggcgacct gagcagacgg gatgtgtctc tgacaatcga
     1201 gaacaccgcc gtgtccgata gcggcgtgta ctgctgtaga gtggaacaca gaggctggtt
     1261 caacgacatg aagatcaccg tgtctctgga aatcgtgccg cctaaagtga ccaccacacc
     1321 tatcgtgacc accgtgccta cagtgacaac cgtgcggaca agcacaacag tgcctaccac
     1381 caccacagtg cccacaacca ctgtgccaac caccatgagc atccccacca caactaccgt
     1441 gctgaccaca atgaccgtgt ccacaacaac cagcgtgccc accactacca gcattcctac
     1501 aaccacatcc gtgcctgtga ccacaaccgt gtctaccttc gtgcctccta tgcctctgcc
     1561 tagacagaat cacgagcctg tggccacctc tcctagtagc cctcagcctg ccgaaacaca
     1621 ccctacaaca ctgcagggcg ccatcagaag agagcctaca agcagccctc tgtacagcta
     1681 caccaccgac ggcaatgata ccgtgaccga gtcctctgac ggcctgtgga acaacaatca
     1741 gacccagctg ttcctggaac actccctgct gaccgccaac accacaaaag gtggcgcgcc
     1801 ggaagatcaa gttgatccaa gattgattga tggcaagggc ggatctctgg aggtgctgtt
     1861 ccagggacca ggcggaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     1921 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     1981 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggta tgcagccctc
     2041 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     2101 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     2161 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     2221 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     2281 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     2341 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     2401 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     2461 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     2521 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     2581 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     2641 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     2701 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2761 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     2821 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     2881 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     2941 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     3001 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     3061 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     3121 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     3181 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     3241 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     3301 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     3361 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     3421 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     3481 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggtggcggag gactgaacga
     3541 catcttcgag gctcagaaaa tcgaatggca cgaacatcac catcaccatc accatcacca
     3601 ttaaggccgg cctgtagtca ctggcacaac gcgttacccg gtaagccaat cgggtataca
     3661 cggtcgtcat actgcagaca gggttcttct actttgcaag atagtcttga gtagtaaaat
     3721 aaatagatag agaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3781 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3841 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     3901 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3961 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4021 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4081 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4141 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4201 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4261 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     4321 agtataatac gacaaggtga ggaactaaac catgaccgag tacaagccca cggtgcgcct
     4381 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     4441 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     4501 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4561 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4621 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4681 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4741 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4801 ggccgagcgc gccggggtgc ccgccttcct ggagacatcc gcgccccgca acctcccctt
     4861 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4921 ctggtgcatg acccgcaagc ccggtgcctg acacgtgcta cgagatttcg attccaccgc
     4981 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5041 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5101 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5161 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5221 gacctctagc tagagcttgg cgtaatcatg gtcattacca atgcttaatc agtgaggcac
     5281 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5341 taactacgat acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac
     5401 cacgctcacc ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5461 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5521 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg
     5581 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5641 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5701 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5761 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5821 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5881 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5941 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6001 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6061 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct
     6121 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6181 ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat taaccaattc
     6241 tgaacattat cgcgagccca tttatacctg aatatggctc ataacacccc ttgctcatga
     6301 ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga
     6361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6481 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6541 gtagttagcc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7021 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     7081 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7141 agcggaaggc gagagtaggg aactgccagg catcaaacta agcagaaggc ccctgacgga
     7201 tggccttttt gcgtttctac aaactctttc tgtgttgtaa aacgacggcc agtcttaagc
     7261 tcgggccccc tgggcggttc tgataacgag taatcgttaa tccgcaaata acgtaaaaac
     7321 ccgcttcggc gggttttttt atggggggag tttagggaaa gagcatttgt cagaatattt
     7381 aagggcgcct gtcactttgc ttgatatatg agaattattt aaccttataa atgagaaaaa
     7441 agcaacgcac tttaaataag atacgttgct ttttcgattg atgaacacct ataattaaac
     7501 tattcatcta ttatttatga ttttttgtat atacaatatt tctagtttgt taaagagaat
     7561 taagaaaata aatctcgaaa ataataaagg gaaaatcagt ttttgatatc aaaattatac
     7621 atgtcaacga taatacaaaa tataatacaa actataagat gttatcagta tttattatca
     7681 tttagaataa attttgtgtc gcccttaatt gtgagcggat aacaattacg agcttcatgc
     7741 acagtggcgt tgactcaata ttggccatta gccatattat tcattggtta tatagcataa
     7801 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     7861 attggctcat gtccaatatg accgccatgt t
//
