LOCUS       sequence-430444-        6871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein 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 6871)
  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 6871)
  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-SIRPB2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6871
                     /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             2447..2491
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2492..2509
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2649..2666)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2655..2766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2884..2904)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2889..3218
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3069..3204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3131..3150
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3266..3313
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3332..3931
                     /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(3332..3351)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3828..3848
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4061..4182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4098..4117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4152..4171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4235..5026,5027..5095))
                     /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     4858..4877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5096..5187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5229..5248
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5254..5273
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5359..5947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5848..5867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6101..6118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6216..6233
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6217..6233
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6689..6705
                     /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 cccagtctag cagaaatgat tggcaggttc tgcagcctga
     1021 gggccctatg cttgtggctg aaggcgaaac cctgctgctg agatgtatgg tcgtgggcag
     1081 ctgcaccgac ggcatgatca aatgggtcaa agtgtctacc caggaccagc aagagatcta
     1141 caacttcaag agaggcagct tccccggcgt gatgcccatg atccagagaa ccagcgagcc
     1201 cctgaactgc gactacagca tctacatcca caacgtgacc agagagcaca ccggcaccta
     1261 ccactgcgtc agatttgatg gcctgagcga gcacagcgag atgaagtctg atgagggcac
     1321 cagcgtgttg gtcaaaggcg ctggcgatcc tgagcctgac ctgtggatta tccagcctca
     1381 agagctggtg ctgggcacaa ccggcgatac cgtgtttctg aattgcaccg tgctcggcga
     1441 cggacctcct ggacctatca gatggtttca aggcgccgga ctgagcagag aggccatcta
     1501 caatttcggc ggcatctctc accccaaaga aacagccgtg caggccagca acaacgactt
     1561 cagcatcctg ctgcagaatg tgtcctctga ggacgccgga acctactact gtgtgaagtt
     1621 ccagcggaag cccaacagac agtatctgag cggccagggc acaagcctga aagtgaaggc
     1681 caagagcacc agcagcaaag aggccgagtt tacaagcgag cctgccaccg aaatgagccc
     1741 caccggtggc gcgccgggcg gcgacaagac ccacacctgc cccccctgcc cagctccaga
     1801 actgctggga gggccttccg tgttcctgtt cccccccaag cccaaggaca ccctgatgat
     1861 cagcaggacc cccgaggtga cctgcgtggt ggtggccgtg tcccacgagg acccagaggt
     1921 gaagttcaac tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcccagaga
     1981 ggagcagtac aacagcacct acagggtggt gtccgtgctg accgtgctgc accaggactg
     2041 gctgaacggc aaagaataca agtgcaaagt ctccaacaag gccctggctg ccccaatcga
     2101 aaagacaatc agcaaggcca agggccagcc acgggagccc caggtgtaca ccctgccccc
     2161 cagccgggag gagatgacca agaaccaggt gtccctgacc tgtctggtga agggcttcta
     2221 ccccagcgat atcgccgtgg agtgggagag caacggccag cccgagaaca actacaagac
     2281 caccccccca gtgctggaca gcgacggcag cttcttcctg tacagcaagc tgaccgtgga
     2341 caagtccagg tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca
     2401 caaccactac acccagaagt ccctgagcct gagccccggc ggcagcggtc ttaatgatat
     2461 tttcgaggct cagaagattg agtggcatga acaccaccac catcaccact aaggccggcc
     2521 tgtagtcact ggcacaacgc gttacccggt aagccaatcg ggtatacacg gtcgtcatac
     2581 tgcagacagg gttcttctac tttgcaagat agtcttgagt agtaaaataa atagatagag
     2641 aaaatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2701 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2761 tgcatcacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2821 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2881 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2941 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3061 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3121 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3181 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3241 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     3301 agtataatac gacaaggtga ggaactaaac catgaccgag tacaagccca cggtgcgcct
     3361 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     3421 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     3481 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     3541 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     3601 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     3661 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     3721 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     3781 ggccgagcgc gccggggtgc ccgccttcct ggagacatcc gcgccccgca acctcccctt
     3841 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     3901 ctggtgcatg acccgcaagc ccggtgcctg acacgtgcta cgagatttcg attccaccgc
     3961 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4021 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4081 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4141 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4201 gacctctagc tagagcttgg cgtaatcatg gtcattacca atgcttaatc agtgaggcac
     4261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4321 taactacgat acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac
     4381 cacgctcacc ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4501 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg
     4561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct
     5101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5161 ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat taaccaattc
     5221 tgaacattat cgcgagccca tttatacctg aatatggctc ataacacccc ttgctcatga
     5281 ccaaaatccc ttaacgtgag ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga
     5341 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5401 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5461 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5521 gtagttagcc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5581 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5641 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5701 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5761 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5821 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5881 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5941 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6001 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6061 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6121 agcggaaggc gagagtaggg aactgccagg catcaaacta agcagaaggc ccctgacgga
     6181 tggccttttt gcgtttctac aaactctttc tgtgttgtaa aacgacggcc agtcttaagc
     6241 tcgggccccc tgggcggttc tgataacgag taatcgttaa tccgcaaata acgtaaaaac
     6301 ccgcttcggc gggttttttt atggggggag tttagggaaa gagcatttgt cagaatattt
     6361 aagggcgcct gtcactttgc ttgatatatg agaattattt aaccttataa atgagaaaaa
     6421 agcaacgcac tttaaataag atacgttgct ttttcgattg atgaacacct ataattaaac
     6481 tattcatcta ttatttatga ttttttgtat atacaatatt tctagtttgt taaagagaat
     6541 taagaaaata aatctcgaaa ataataaagg gaaaatcagt ttttgatatc aaaattatac
     6601 atgtcaacga taatacaaaa tataatacaa actataagat gttatcagta tttattatca
     6661 tttagaataa attttgtgtc gcccttaatt gtgagcggat aacaattacg agcttcatgc
     6721 acagtggcgt tgactcaata ttggccatta gccatattat tcattggtta tatagcataa
     6781 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
     6841 attggctcat gtccaatatg accgccatgt t
//
