LOCUS       sequence-385764-        8329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            COMP5AP-AviTag-9xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8329)
  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 8329)
  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-PSG9-COMP5AP-AviTag-9xHis
FEATURES             Location/Qualifiers
     source          1..8329
                     /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             2168..2203
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             2213..2236
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3896..3940
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3941..3967
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(4107..4124)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4113..4224
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4342..4362)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4347..4676
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4527..4662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4589..4608
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4724..4771
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4790..5389
                     /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(4790..4809)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5286..5306
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5519..5640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5556..5575)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5610..5629
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5693..6484,6485..6553))
                     /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     6316..6335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6554..6645)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6687..6706
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6712..6731
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6817..7405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7306..7325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7559..7576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7674..7691
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7675..7691
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8147..8163
                     /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 ccgaagtgac aattgaagcc cagcctccta aggtgtccga
     1021 gggcaaagat gtgctgctgc tggtgcacaa cctgcctcag aatctgcccg gctacttctg
     1081 gtacaagggc gagatgaccg acctgtacca ctacatcatc agctacatcg tggacggcaa
     1141 gatcatcatc tacggccctg cctacagcgg cagagaaacc gtgtacagca atgccagcct
     1201 gctgatccag aacgtgacca gaaaggacgc cggcacctac acactgcaca tcatcaagag
     1261 aggcgacgag acacgggaag agatccggca cttcaccttc acactgtacc tggaaacccc
     1321 taagccttac atcagcagct ccaatctgaa ccccagagaa gccatggaag ccgtgcggct
     1381 gatctgcgat cctgagacac tggatgccag ctacctgtgg tggatgaatg gccagtctct
     1441 gcccgtgaca cacagactgc agctgagcaa gaccaacaga accctgtacc tgttcggcgt
     1501 gaccaagtat atcgctggcc cctacgagtg cgagatcaga aatcctgtgt ccgccagcag
     1561 atctgacccc gtgactctga atctgctgcc caagctgccc attccgtaca tcaccatcaa
     1621 caacctgaat ccgcgcgaga acaaggacgt gctggccttt acctgcgagc ccaagagcga
     1681 gaactacacc tacatctggt ggctcaacgg ccagagcctt cctgtgtctc ctggcgtgaa
     1741 gaggcccatc gagaacagaa tcctgatcct gcctagcgtg acccggaacg agacaggccc
     1801 ttaccagtgc gaaatccggg atagatacgg cggcctgaga agcaaccctg tgatcctgaa
     1861 tgtgctgtac ggccccgacc tgccaagaat ctaccccagc ttcacctact acagaagcgg
     1921 cgagaacctg gacctgagct gcttcacaga gagcaaccct ccagccgagt acttctggac
     1981 catcaacggc aagtttcagc agagcggcca gaagctgttc attcctcaga tcaccagaaa
     2041 ccacagcggc ctgtacgcct gctctgtgca taatagcgcc accggcaaag aaatcagcaa
     2101 gagcatgacc gtgaaagtgt ctggcccttg ccacggcgat ctgaccgaat ctcaatctgg
     2161 cgcgccggaa gatcaagttg atccaagatt gattgatggc aagggcggat ctctggaggt
     2221 gctgttccag ggaccaggcg gaagcggcaa tcccggcggc gatctggccc cccagatgct
     2281 gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg
     2341 cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggtatgca
     2401 gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa
     2461 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
     2521 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
     2581 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
     2641 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
     2701 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
     2761 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
     2821 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
     2881 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
     2941 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
     3001 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
     3061 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
     3121 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
     3181 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
     3241 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
     3301 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
     3361 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
     3421 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
     3481 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
     3541 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
     3601 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
     3661 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
     3721 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
     3781 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
     3841 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtg gcggaggact
     3901 gaacgacatc ttcgaggctc agaaaatcga atggcacgaa catcaccatc accatcacca
     3961 tcaccattaa ggccggcctg tagtcactgg cacaacgcgt tacccggtaa gccaatcggg
     4021 tatacacggt cgtcatactg cagacagggt tcttctactt tgcaagatag tcttgagtag
     4081 taaaataaat agatagagaa aatcagcctc gactgtgcct tctagttgcc agccatctgt
     4141 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4201 ctaataaaat gaggaaattg catcacaaca ctcaacccta tctcggtcta ttcttttgat
     4261 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4321 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4381 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4561 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4621 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4681 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     4741 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tgaccgagta
     4801 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4861 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4921 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4981 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5041 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5101 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5161 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5221 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacatccgc
     5281 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5341 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac acgtgctacg
     5401 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5461 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5521 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5581 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5641 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt cattaccaat
     5701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagcgctg
     5821 cgatgatacc gcgagaacca cgctcaccgg ctccggattt atcagcaata aaccagccag
     5881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6001 ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6541 gttgaatact catattcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gtcagtgtta
     6661 caaccaatta accaattctg aacattatcg cgagcccatt tatacctgaa tatggctcat
     6721 aacacccctt gctcatgacc aaaatccctt aacgtgagtt acgcgcgcgt cgttccactg
     6781 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6841 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6901 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6961 tgttcttcta gtgtagccgt agttagccca ccacttcaag aactctgtag caccgcctac
     7021 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7081 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7141 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7201 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7261 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7321 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7381 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7441 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7501 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7561 cgagtcagtg agcgaggaag cggaaggcga gagtagggaa ctgccaggca tcaaactaag
     7621 cagaaggccc ctgacggatg gcctttttgc gtttctacaa actctttctg tgttgtaaaa
     7681 cgacggccag tcttaagctc gggccccctg ggcggttctg ataacgagta atcgttaatc
     7741 cgcaaataac gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt tagggaaaga
     7801 gcatttgtca gaatatttaa gggcgcctgt cactttgctt gatatatgag aattatttaa
     7861 ccttataaat gagaaaaaag caacgcactt taaataagat acgttgcttt ttcgattgat
     7921 gaacacctat aattaaacta ttcatctatt atttatgatt ttttgtatat acaatatttc
     7981 tagtttgtta aagagaatta agaaaataaa tctcgaaaat aataaaggga aaatcagttt
     8041 ttgatatcaa aattatacat gtcaacgata atacaaaata taatacaaac tataagatgt
     8101 tatcagtatt tattatcatt tagaataaat tttgtgtcgc ccttaattgt gagcggataa
     8161 caattacgag cttcatgcac agtggcgttg actcaatatt ggccattagc catattattc
     8221 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat
     8281 cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgtt
//
