LOCUS       sequence-401036-        7300 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 7300)
  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 7300)
  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-NTRK3-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7300
                     /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             2876..2920
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2921..2938
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3078..3095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3084..3195
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3313..3333)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3318..3647
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3498..3633
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3560..3579
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3695..3742
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3761..4360
                     /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(3761..3780)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4257..4277
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4490..4611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4527..4546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4581..4600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4664..5455,5456..5524))
                     /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     5287..5306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5525..5616)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5658..5677
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5683..5702
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5788..6376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6277..6296
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6530..6547
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6645..6662
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6646..6662
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7118..7134
                     /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 cctgtcctgc taattgcgtg tgtagcaaga ccgagatcaa
     1021 ctgcagaagg cccgacgacg gcaatctgtt ccctctgctg gaaggccagg acagcggcaa
     1081 ctctaatggc aacgccagca tcaacatcac cgacatcagc cggaacatca cctccatcca
     1141 catcgagaac tggcggagcc tgcacaccct gaacgccgtg gatatggaac tgtacaccgg
     1201 cctgcagaag ctgaccatca agaatagcgg cctgcggagc atccagccta gagcctttgc
     1261 caagaatccc catctgcggt acatcaacct gagcagcaac cggctgacca cactgagctg
     1321 gcagctgttt cagaccctga gcctgagaga actgcagctg gaacagaact tcttcaactg
     1381 ctcctgcgac atccggtgga tgcagctgtg gcaagaacag ggcgaagcca agctgaacag
     1441 ccagaacctg tactgcatca acgccgacgg ctctcagctg cccctgttca gaatgaacat
     1501 cagccagtgc gacctgcctg agatcagcgt gtcccacgtg aacctgaccg tcagagaagg
     1561 cgacaacgcc gtgatcacct gtaacggctc tggcagccct ctgcctgatg tggattggat
     1621 cgtgaccgga ctgcagagca tcaataccca ccagaccaac ctgaactgga ccaacgtgca
     1681 cgccatcaat ctgaccctgg tcaacgtgac ctccgaggac aatggcttca ccctgacctg
     1741 tatcgccgag aacgtcgtgg gcatgagcaa tgcctctgtg gccctgaccg tgtactaccc
     1801 tcctagagtg gtgtccctgg aagaacccga gctgagactg gaacactgca tcgagtttgt
     1861 cgtgcggggc aaccctcctc ctacactgca ttggctgcac aacggccagc ctctgagaga
     1921 gtccaagatc atccacgtcg agtactacca agagggcgag atctccgagg gctgtctgct
     1981 gttcaacaag cccacacact acaacaacgg caactacacc ctgatcgcta agaaccctct
     2041 gggcaccgcc aaccagacaa tcaacggcca cttcctgaaa gagccctttc cagagagcac
     2101 cgacaacttc atcctgttcg acgaggtgtc ccctacacct cctatcaccg tgacacacaa
     2161 gcccgaagag gacacaggcg cgccgggcgg cgacaagacc cacacctgcc ccccctgccc
     2221 agctccagaa ctgctgggag ggccttccgt gttcctgttc ccccccaagc ccaaggacac
     2281 cctgatgatc agcaggaccc ccgaggtgac ctgcgtggtg gtggccgtgt cccacgagga
     2341 cccagaggtg aagttcaact ggtacgtgga cggcgtggag gtgcacaacg ccaagaccaa
     2401 gcccagagag gagcagtaca acagcaccta cagggtggtg tccgtgctga ccgtgctgca
     2461 ccaggactgg ctgaacggca aagaatacaa gtgcaaagtc tccaacaagg ccctggctgc
     2521 cccaatcgaa aagacaatca gcaaggccaa gggccagcca cgggagcccc aggtgtacac
     2581 cctgcccccc agccgggagg agatgaccaa gaaccaggtg tccctgacct gtctggtgaa
     2641 gggcttctac cccagcgata tcgccgtgga gtgggagagc aacggccagc ccgagaacaa
     2701 ctacaagacc acccccccag tgctggacag cgacggcagc ttcttcctgt acagcaagct
     2761 gaccgtggac aagtccaggt ggcagcaggg caacgtgttc agctgcagcg tgatgcacga
     2821 ggccctgcac aaccactaca cccagaagtc cctgagcctg agccccggcg gcagcggtct
     2881 taatgatatt ttcgaggctc agaagattga gtggcatgaa caccaccacc atcaccacta
     2941 aggccggcct gtagtcactg gcacaacgcg ttacccggta agccaatcgg gtatacacgg
     3001 tcgtcatact gcagacaggg ttcttctact ttgcaagata gtcttgagta gtaaaataaa
     3061 tagatagaga aaatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3121 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3181 tgaggaaatt gcatcacaac actcaaccct atctcggtct attcttttga tttataaggg
     3241 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3301 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3361 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3421 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3481 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3541 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3601 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3661 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     3721 tatcggcata gtataatacg acaaggtgag gaactaaacc atgaccgagt acaagcccac
     3781 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     3841 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     3901 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     3961 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4021 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4081 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4141 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4201 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacatccg cgccccgcaa
     4261 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4321 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cacgtgctac gagatttcga
     4381 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4441 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4501 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4561 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4621 tataccgtcg acctctagct agagcttggc gtaatcatgg tcattaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagcgct gcgatgatac
     4801 cgcgagaacc acgctcaccg gctccggatt tatcagcaat aaaccagcca gccggaaggg
     4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccatcgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcatattctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggtcagtgtt acaaccaatt
     5641 aaccaattct gaacattatc gcgagcccat ttatacctga atatggctca taacacccct
     5701 tgctcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
     5761 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5821 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5881 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5941 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
     6001 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6061 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6181 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6421 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6481 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6541 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     6601 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     6661 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     6721 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     6781 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     6841 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     6901 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     6961 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     7021 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     7081 ttattatcat ttagaataaa ttttgtgtcg cccttaattg tgagcggata acaattacga
     7141 gcttcatgca cagtggcgtt gactcaatat tggccattag ccatattatt cattggttat
     7201 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     7261 tacatttata ttggctcatg tccaatatga ccgccatgtt
//
