LOCUS       sequence-444426-        8644 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 8644)
  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 8644)
  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-MAG-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     149..168
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      254..842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     996..1013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1111..1128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1112..1128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1584..1600
                     /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)."
     enhancer        1767..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2347..2371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2581..2599
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2581..2598
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2612..2631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4249..4284
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             4294..4317
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5977..6021
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6022..6048
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(6188..6205)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6194..6305
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6423..6443)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6428..6757
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6608..6743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6670..6689
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6805..6852
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6871..7470
                     /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(6871..6890)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7367..7387
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7600..7721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7637..7656)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7691..7710
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7774..8565,8566..8634))
                     /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"
     promoter        complement(join(8635..8644,1..82))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
      121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
      181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
      241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
     1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
     1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
     1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
     1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
     1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
     1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
     1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
     1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
     1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
     1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
     1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
     2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
     2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
     2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgccgg acattgggga
     2761 gcttggatgc cttctagcat cagcgccttc gagggaacct gtgtgtcaat cccctgcaga
     2821 ttcgacttcc ccgacgaact gaggcctgct gtggttcacg gcgtgtggta cttcaacagc
     2881 ccctatccta agaactaccc tcctgtggtg ttcaagagca gaacccaggt ggtgcacgag
     2941 agcttccagg gcagaagtag actgctgggc gatctgggcc tgagaaactg tacactgctg
     3001 ctgagcaacg tgtcccctga actcggcggc aagtactact tcagaggcga tctcggcggc
     3061 tacaaccagt acacctttag cgagcacagc gtgctggaca tcgtgaacac ccctaacatc
     3121 gtggtgcctc cagaagtggt ggccggcaca gaggtggaag tgtcctgtat ggtgcccgac
     3181 aactgccctg agctgaggcc agaactgtct tggctgggac acgaaggact gggagaacct
     3241 gctgttctgg gcagactgag agaggacgaa ggcacctggg tgcaagtgtc cctgctgcac
     3301 tttgtgccta ccagagaggc caacggccac agactgggtt gtcaggccag ctttcccaac
     3361 accacactgc agtttgaggg ctacgccagc atggacgtga agtaccctcc agtgatcgtg
     3421 gaaatgaaca gcagcgtgga agccatcgag ggctctcatg tgtctctgct gtgtggcgcc
     3481 gacagcaatc ctcctcctct gctgacctgg atgagagatg gcaccgtgct gagagaagcc
     3541 gtggccgaat ctctgctgct ggaactggaa gaagtgaccc ctgccgagga tggcgtgtac
     3601 gcttgtctgg ccgagaatgc ctacggccag gacaatagaa ccgtgggcct gtccgtgatg
     3661 tacgcccctt ggaagcctac cgtgaacggc acaatggtgg ccgtggaagg cgagacagtg
     3721 tccatcctgt gtagcaccca gagcaacccc gatcctatcc tgaccatctt caaagagaag
     3781 cagatcctga gcaccgtgat ctacgagagc gaactgcagc tcgaactgcc cgctgtgtcc
     3841 ccagaggatg atggcgaata ttggtgcgtg gcagagaacc agtacggcca gagagccacc
     3901 gccttcaacc tgagcgtgga atttgctccc gtgctgctgc tcgagagcca ttgtgctgcc
     3961 gccagagata ccgtgcagtg cctgtgtgtg gtcaagtcta accccgagcc tagcgtggcc
     4021 tttgagctgc ccagcagaaa cgtgaccgtg aatgagagcg agcgcgagtt cgtgtacagc
     4081 gagagatctg gactggtgct gaccagcatc ctgacactga gaggacaggc tcaggcccct
     4141 cctagagtga tctgcaccgc cagaaatctg tacggcgcca agagcctgga actgccattt
     4201 cagggcgccc acagactcat gtgggccaag attggacctg gcgcgccgga agatcaagtt
     4261 gatccaagat tgattgatgg caagggcgga tctctggagg tgctgttcca gggaccaggc
     4321 ggaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct gcaggagacc
     4381 aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga gatcaccttc
     4441 ctgaagaaca ccgtgatgga gtgcgacgcc tgcggtatgc agccctctag aggctccggc
     4501 tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc agccgaggcc
     4561 ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct catcatcttc
     4621 ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa agggcagaag
     4681 aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata tgtggctctg
     4741 tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc cacggcctac
     4801 ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc ccgctttaac
     4861 cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc caagaaagca
     4921 gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc agccggcacc
     4981 tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc ctcggcccgc
     5041 caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat tgacgtgatc
     5101 ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga gtacccagat
     5161 gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga atggctggcg
     5221 aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc ttccctggac
     5281 ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata cgagatccac
     5341 cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct gcgcctgctg
     5401 agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga ccatggtcat
     5461 catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga cgccattgag
     5521 agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc cgaccactcc
     5581 cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg gctggcccct
     5641 ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg tccaggctat
     5701 gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag ccccgagtat
     5761 cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga cgtggcggtg
     5821 ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac cttcatagcg
     5881 cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct ggcgcccccc
     5941 gccggcacca ccgacgccgc gcacccgggt ggcggaggac tgaacgacat cttcgaggct
     6001 cagaaaatcg aatggcacga acatcaccat caccatcacc atcaccatta aggccggcct
     6061 gtagtcactg gcacaacgcg ttacccggta agccaatcgg gtatacacgg tcgtcatact
     6121 gcagacaggg ttcttctact ttgcaagata gtcttgagta gtaaaataaa tagatagaga
     6181 aaatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6241 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6301 gcatcacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     6361 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     6421 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6481 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     6541 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     6601 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6661 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     6721 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     6781 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     6841 gtataatacg acaaggtgag gaactaaacc atgaccgagt acaagcccac ggtgcgcctc
     6901 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     6961 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     7021 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     7081 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     7141 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     7201 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     7261 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     7321 gccgagcgcg ccggggtgcc cgccttcctg gagacatccg cgccccgcaa cctccccttc
     7381 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     7441 tggtgcatga cccgcaagcc cggtgcctga cacgtgctac gagatttcga ttccaccgcc
     7501 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     7561 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     7621 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7681 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     7741 acctctagct agagcttggc gtaatcatgg tcattaccaa tgcttaatca gtgaggcacc
     7801 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7861 aactacgata cgggagggct taccatctgg ccccagcgct gcgatgatac cgcgagaacc
     7921 acgctcaccg gctccggatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7981 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8041 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccatcgcta caggcatcgt
     8101 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8161 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8221 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8281 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8341 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8401 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8461 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8521 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8581 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatattctt
     8641 cctt
//
