LOCUS       sequence-368825-        6469 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 6469)
  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 6469)
  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-MFAP3L-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(140..187)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(235..564)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      249..384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(303..322)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     549..569
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    687..798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     787..804
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(944..961)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(962..1006)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2186..2205)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2218..2236)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2219..2236)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2446..2470)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2467..2670)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2496..2516)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2671..3050
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3217..3233
                     /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)."
     primer_bind     complement(3689..3706)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3689..3705)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3804..3821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3975..4563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4055..4074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4649..4668)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4674..4693)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4735..4826
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4827..4895,4896..5687)
                     /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     complement(5045..5064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5740..5861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5751..5770)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5805..5824
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5991..6469,1..121))
                     /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(6074..6094)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
       61 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
      121 tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat
      181 tgtcaacacg tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca
      241 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc
      301 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
      361 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      421 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      481 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      541 ctttccacac cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa
      601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      661 tagaccgaga tagggttgag tgttgtgatg caatttcctc attttattag gaaaggacag
      721 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
      781 ggcaactaga aggcacagtc gaggctgatt ttctctatct atttatttta ctactcaaga
      841 ctatcttgca aagtagaaga accctgtctg cagtatgacg accgtgtata cccgattggc
      901 ttaccgggta acgcgttgtg ccagtgacta caggccggcc ttagtggtga tggtggtggt
      961 gttcatgcca ctcaatcttc tgagcctcga aaatatcatt aagaccgctg ccgccggggc
     1021 tcaggctcag ggacttctgg gtgtagtggt tgtgcagggc ctcgtgcatc acgctgcagc
     1081 tgaacacgtt gccctgctgc cacctggact tgtccacggt cagcttgctg tacaggaaga
     1141 agctgccgtc gctgtccagc actggggggg tggtcttgta gttgttctcg ggctggccgt
     1201 tgctctccca ctccacggcg atatcgctgg ggtagaagcc cttcaccaga caggtcaggg
     1261 acacctggtt cttggtcatc tcctcccggc tggggggcag ggtgtacacc tggggctccc
     1321 gtggctggcc cttggccttg ctgattgtct tttcgattgg ggcagccagg gccttgttgg
     1381 agactttgca cttgtattct ttgccgttca gccagtcctg gtgcagcacg gtcagcacgg
     1441 acaccaccct gtaggtgctg ttgtactgct cctctctggg cttggtcttg gcgttgtgca
     1501 cctccacgcc gtccacgtac cagttgaact tcacctctgg gtcctcgtgg gacacggcca
     1561 ccaccacgca ggtcacctcg ggggtcctgc tgatcatcag ggtgtccttg ggcttggggg
     1621 ggaacaggaa cacggaaggc cctcccagca gttctggagc tgggcagggg gggcaggtgt
     1681 gggtcttgtc gccgcccggc gcgcccatgt cgccgctggt gaagatcact ctcagtgtca
     1741 cggtgttgtt cacggtgccg tagatattgc tggccacgca ggtatacttg ccccggtcgc
     1801 tgaaggacac cttggtgatg ttcagcaggc cagaatcgtg catctgccac ttgcctccgc
     1861 cgcgttcttt ctcgtcctct tcctctttca gcagcttgcc gatgctgttg taccacttga
     1921 actgaggatc ggggatgccg tacacgctgc agttgatcag ggcgctgttg ccctctttca
     1981 cgatgatgtg gtcggttctg gcaatgatca cgggcacgct gcccagcacc acattggtgc
     2041 cattcagggt gctattggtc acgctcttgg cggccgcagc gaacaccagg cagaagatgt
     2101 agctcagggc aatgatggtc ttcatggtgg cgctagcagg cgcgttttcg atagaaccga
     2161 ggtgcagttg gacctgggag tggacacctg tggagagaaa ggcaaagtgg atgttattct
     2221 atagtgtcac ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg
     2281 ttctaacgaa gccaaggggg tgggcctata gactctatag gcggtactta cgtcactctt
     2341 ggcacgggga atccgcgttc caatgcaccg ttcccggccg cggaggctgg atcggtcccg
     2401 gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac
     2461 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     2521 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     2581 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     2641 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     2701 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     2761 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     2821 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     2881 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     2941 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3001 taatgacccc gtaattgatt actattaata actagtcaat aatcaatgcc aacatggcgg
     3061 tcatattgga catgagccaa tataaatgta catattatga tatagataca acgtatgcaa
     3121 tggccaatag ccaatattga tttatgctat ataaccaatg aataatatgg ctaatggcca
     3181 atattgagtc aacgccactg tgcatgaagc tcgtaattgt tatccgctca caattaaggg
     3241 cgacacaaaa tttattctaa atgataataa atactgataa catcttatag tttgtattat
     3301 attttgtatt atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta
     3361 ttttcgagat ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa
     3421 atcataaata atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta
     3481 tcttatttaa agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca
     3541 agcaaagtga caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca
     3601 taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc
     3661 agaaccgccc agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt
     3721 tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt
     3781 tccctactct cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3841 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3901 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4141 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4381 ttgaagtggt gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4441 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4561 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta
     4621 actcacgtta agggattttg gtcatgagca aggggtgtta tgagccatat tcaggtataa
     4681 atgggctcgc gataatgttc agaattggtt aattggttgt aacactgacc cctatttgtt
     4741 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4801 ttcaataata ttgaaaaagg aagaatatga gtattcaaca tttccgtgtc gcccttattc
     4861 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4921 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4981 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5041 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5101 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5161 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5221 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5281 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5341 caaacgacga gcgtgacacc acgatgcctg tagcgatggc aacaacgttg cgcaaactat
     5401 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5461 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5521 aatccggagc cggtgagcgt ggttctcgcg gtatcatcgc agcgctgggg ccagatggta
     5581 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5641 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaatga ccatgattac
     5701 gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg
     5761 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     5821 ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc
     5881 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     5941 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtagcacgtg tcaggcaccg
     6001 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg
     6061 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggatgtctc caggaaggcg
     6121 ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc
     6181 ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc
     6241 cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac
     6301 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg
     6361 gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg
     6421 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggc
//
