LOCUS       sequence-303149-        9064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein partial extracellular 
            domain fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9064)
  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 9064)
  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-IGSF10(Trunc2)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..9064
                     /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             6406..6450
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6451..6468
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6608..6625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6614..6725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6843..6863)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6848..7177
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7028..7163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7090..7109
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7225..7272
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7291..7890
                     /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(7291..7310)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7787..7807
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8020..8141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8057..8076)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8111..8130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8194..8985,8986..9054))
                     /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     8817..8836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9055..9064,1..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 cggccgccgt gaaaatgaag
     2761 ggccaaagac ctctggaaca cgacggcgag acagaaggct ctggactgga cgagagcaac
     2821 cctatcgctc acctgaaaga gcctccaggc gcacagctga gaacaagcgc tctgatggaa
     2881 gccgaagtgg gcaagcacac cagcagcacc agcaagcggc acaactacag agagctgacc
     2941 ctgcagagaa gaggcgacag cacccacaga cggttcagag agaacagacg gcacttccca
     3001 cctagcgcca gaagaatcga tccccagcat tgggccgctc tgctggaaaa ggccaagaaa
     3061 aacgccatgc ctgacaagcg cgagaacacc accgtttctc cacctccagt ggtcacccag
     3121 ctgcctaaca ttcccggcga agaggatgac agctctggca tgctggccct gcacgaggaa
     3181 ttcatggtgc ctgccacaaa ggccctgaac ctgcctgcca gaacagtgac cgccgacagc
     3241 agaaccatca gcgacagccc catgaccaac atcaactacg gcaccgagtt cagccccgtg
     3301 gtcaactctc agatcctgcc tcctgaggaa cccaccgact tcaagctgag caccgccatc
     3361 aagaccaccg ccatgagcaa gaacatcaac cccaccatga gcagccagat ccagggcacc
     3421 acaaaccagc acagcagcac agtgttccct ctgctgctgg gcgccaccga gtttcaggat
     3481 agcgatcaga tgggcagagg cagagagcac ttccagagca gacctccaat caccgtgcgg
     3541 accatgatca aggacgtgaa cgtgaagatg ctgtccagca ccaccaacaa actgctgctc
     3601 gagtccgtga ataccaccaa tagccaccag accagcgtgc gggaagtgtc cgagcctaga
     3661 cacaaccact tctacagcca caccacacag atcctgtcca ccagcacatt ccccagcgat
     3721 cctcacacag ccgctcactc tcagttcccc attcctcgga actctaccgt gaacatccct
     3781 ctgttcagaa gattcggccg gcagagaaag atcggcggca ggggcagaat catctcccca
     3841 tacagaaccc ctgtgctgcg gcggcaccgg tactccatct ttagaagcac cacacggggc
     3901 agcagcgaga agtctaccac agcctttagc gccaccgtgc tgaacgtgac ctgtctgagc
     3961 tgtctgccca gagagagact gaccacagcc acagctgccc tgagctttcc atctgccgct
     4021 cctatcacct ttcctaaggc cgatatcgcc agagtgccca gcgaggaatc tacaaccctg
     4081 gtgcagaatc cactgctgct gcttgagaac aaacccagcg tggaaaagac cacacctacc
     4141 atcaagtact tccggaccga gatcagccaa gtgaccccaa caggcgccgt gatgacatac
     4201 gcccctacca gcattcccat ggaaaaaacc cacaaagtga acgccagcta tccccgggtg
     4261 tcctccacaa atgaggccaa gagggacagc gtgatcacct ctagtctgag cggcgccatc
     4321 accaagcctc caatgaccat cattgccatc acacggttca gccggcggaa gatcccttgg
     4381 cagcagaact tcgtgaacaa tcacaacccc aagggcagac tgcggaatca gcacaaggtg
     4441 tccctgcaga aaagcacagc cgtgatgctg cccaagacaa gccctgctct gcctcgggac
     4501 aaagtgtccc ctttccactt caccacactg agcaccagcg tgatgcagat ccctagcaac
     4561 accctgacca ccgctcacca caccaccaca aagacccaca atcctggcag cctgcctacc
     4621 aagaaagagc tgccctttcc tcctctgaac cccatgctgc cctccatcat cagcaaggac
     4681 agctccacca agagcatcat ctctacccag accgccattc ctgccaccac tcctacattt
     4741 cccgcctccg tgattaccta cgagacacag accgagcgga gcagagccca gaccattcag
     4801 cgggaacaag agccccagaa gaagaaccgg accgatccta acatcagccc cgatcagtcc
     4861 agcggcttca caacacccac agccatgaca ccacctgtgc tgacaacagc cgagacaagc
     4921 gtgaagccta gcgtgtccgc cttcacacac agccctccag agaacacaac cggcatctct
     4981 agcaccatca gcttccacag caggaccctg aatctgaccg acgtgatcga ggaactggcc
     5041 caggcctcta cacagaccct gaagtctaca atcgccagcg agacaaccct gagcagcaag
     5101 agccaccagt ccaccacaac cagaaaggcc atcatccggc actctacaat ccctccattc
     5161 ctgtccagct ccgccacact gatgcccgtg cctatcagcc cacctttcac acagagagcc
     5221 gtgaccgaca acgtggccac acctatctct ggcctgatga ccaataccgt cgtgaagctg
     5281 cacgagagca gccggcacaa tgccaaacct cagcagctgg tggcagaggt ggccacatct
     5341 cctaaggtgc accccaacgc caagttcacc atcggcacca cacacttcat ctacagcaac
     5401 ctgctgcaca gcacccctat gcctgctctg acaaccgtga agtcccagaa cagcaagctg
     5461 accccatctc cttgggccga gaaccagttc tggcacaagc cctatagcga gatcgccgag
     5521 aagggcaaga aacccgaggt gtcaatgctg gccacaacag gactgagcga ggccacaact
     5581 ctggtgtccg attgggacgg acagaagaac accaagaagt ccgacttcga caaaaagccc
     5641 gtgcaagagg ccactacctc caagctgctg cctttcgata gcctgagccg gtacatcttc
     5701 gagaaaggcg cgccgggcgg cgacaagacc cacacctgcc ccccctgccc agctccagaa
     5761 ctgctgggag ggccttccgt gttcctgttc ccccccaagc ccaaggacac cctgatgatc
     5821 agcaggaccc ccgaggtgac ctgcgtggtg gtggccgtgt cccacgagga cccagaggtg
     5881 aagttcaact ggtacgtgga cggcgtggag gtgcacaacg ccaagaccaa gcccagagag
     5941 gagcagtaca acagcaccta cagggtggtg tccgtgctga ccgtgctgca ccaggactgg
     6001 ctgaacggca aagaatacaa gtgcaaagtc tccaacaagg ccctggctgc cccaatcgaa
     6061 aagacaatca gcaaggccaa gggccagcca cgggagcccc aggtgtacac cctgcccccc
     6121 agccgggagg agatgaccaa gaaccaggtg tccctgacct gtctggtgaa gggcttctac
     6181 cccagcgata tcgccgtgga gtgggagagc aacggccagc ccgagaacaa ctacaagacc
     6241 acccccccag tgctggacag cgacggcagc ttcttcctgt acagcaagct gaccgtggac
     6301 aagtccaggt ggcagcaggg caacgtgttc agctgcagcg tgatgcacga ggccctgcac
     6361 aaccactaca cccagaagtc cctgagcctg agccccggcg gcagcggtct taatgatatt
     6421 ttcgaggctc agaagattga gtggcatgaa caccaccacc atcaccacta aggccggcct
     6481 gtagtcactg gcacaacgcg ttacccggta agccaatcgg gtatacacgg tcgtcatact
     6541 gcagacaggg ttcttctact ttgcaagata gtcttgagta gtaaaataaa tagatagaga
     6601 aaatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6661 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6721 gcatcacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     6781 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     6841 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6901 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     6961 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     7021 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     7081 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     7141 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     7201 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     7261 gtataatacg acaaggtgag gaactaaacc atgaccgagt acaagcccac ggtgcgcctc
     7321 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     7381 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     7441 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     7501 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     7561 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     7621 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     7681 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     7741 gccgagcgcg ccggggtgcc cgccttcctg gagacatccg cgccccgcaa cctccccttc
     7801 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     7861 tggtgcatga cccgcaagcc cggtgcctga cacgtgctac gagatttcga ttccaccgcc
     7921 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     7981 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     8041 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8101 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     8161 acctctagct agagcttggc gtaatcatgg tcattaccaa tgcttaatca gtgaggcacc
     8221 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8281 aactacgata cgggagggct taccatctgg ccccagcgct gcgatgatac cgcgagaacc
     8341 acgctcaccg gctccggatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8401 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8461 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccatcgcta caggcatcgt
     8521 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8581 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8641 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8701 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8761 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8821 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8881 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8941 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9001 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatattctt
     9061 cctt
//
