LOCUS       sequence-386092-        7066 bp    DNA     circular SYN 15-APR-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 7066)
  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 7066)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-HEPACAM2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7066
                     /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             2642..2686
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2687..2704
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2844..2861)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2850..2961
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3079..3099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3084..3413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3264..3399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3326..3345
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3461..3508
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3527..4126
                     /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(3527..3546)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4023..4043
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4256..4377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4293..4312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4347..4366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4430..5221,5222..5290))
                     /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     5053..5072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5291..5382)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5424..5443
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5449..5468
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5554..6142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6043..6062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6296..6313
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6411..6428
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6412..6428
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6884..6900
                     /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 ccttgaaagt gacagtgcct tctcatacag tgcacggcgt
     1021 tagaggacag gccctgtacc tgcctgtgca ctacggattt cacacccctg ccagcgacat
     1081 ccagatcatc tggctgttcg agaggcccca caccatgcct aagtatctgc tgggcagcgt
     1141 gaacaagagc gtggtgcctg atctggaata ccagcacaag ttcaccatga tgcctccaaa
     1201 cgccagcctg ctgatcaacc ctctgcagtt ccccgacgag ggcaactaca tcgtgaaagt
     1261 gaacatccaa ggcaacggca ccctgagcgc cagccagaaa attcaagtga ccgtggacga
     1321 ccccgtgacc aagcctgtgg ttcagattca tcctcctagc ggagccgtgg aatacgtggg
     1381 caatatgacc ctgacctgtc acgtggaagg cggcacaaga ctggcctacc agtggctgaa
     1441 aaacggcaga cctgtgcaca ccagcagcac ctacagcttc agccctcaga acaacaccct
     1501 gcacattgcc cctgtgacca aagaggacat cggcaattac agctgcctcg tgcggaaccc
     1561 cgtgtccgag atggaaagcg acatcatcat gcccatcatc tactacggcc cctacggcct
     1621 gcaagtgaac agcgataagg gcctgaaagt gggcgaagtg ttcaccgtgg atctgggcga
     1681 agccatcctg ttcgattgca gcgccgactc tcaccctcct aatacctact cctggatcag
     1741 acggaccgac aacaccacct acatcatcaa gcacggcccc agactggaag tggcctctga
     1801 gaaagtggcc cagaaaacca tggactacgt gtgctgcgcc tacaacaaca tcaccggcag
     1861 acaggacgag acacacttca ccgtgatcat cacaagcgtg ggcctcgaga agctggctca
     1921 gaagggaaag agcctgtctc ctggcgcgcc gggcggcgac aagacccaca cctgcccccc
     1981 ctgcccagct ccagaactgc tgggagggcc ttccgtgttc ctgttccccc ccaagcccaa
     2041 ggacaccctg atgatcagca ggacccccga ggtgacctgc gtggtggtgg ccgtgtccca
     2101 cgaggaccca gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa
     2161 gaccaagccc agagaggagc agtacaacag cacctacagg gtggtgtccg tgctgaccgt
     2221 gctgcaccag gactggctga acggcaaaga atacaagtgc aaagtctcca acaaggccct
     2281 ggctgcccca atcgaaaaga caatcagcaa ggccaagggc cagccacggg agccccaggt
     2341 gtacaccctg ccccccagcc gggaggagat gaccaagaac caggtgtccc tgacctgtct
     2401 ggtgaagggc ttctacccca gcgatatcgc cgtggagtgg gagagcaacg gccagcccga
     2461 gaacaactac aagaccaccc ccccagtgct ggacagcgac ggcagcttct tcctgtacag
     2521 caagctgacc gtggacaagt ccaggtggca gcagggcaac gtgttcagct gcagcgtgat
     2581 gcacgaggcc ctgcacaacc actacaccca gaagtccctg agcctgagcc ccggcggcag
     2641 cggtcttaat gatattttcg aggctcagaa gattgagtgg catgaacacc accaccatca
     2701 ccactaaggc cggcctgtag tcactggcac aacgcgttac ccggtaagcc aatcgggtat
     2761 acacggtcgt catactgcag acagggttct tctactttgc aagatagtct tgagtagtaa
     2821 aataaataga tagagaaaat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2881 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2941 ataaaatgag gaaattgcat cacaacactc aaccctatct cggtctattc ttttgattta
     3001 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3061 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3421 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     3481 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatga ccgagtacaa
     3541 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     3601 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     3661 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     3721 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     3781 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     3841 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     3901 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     3961 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga catccgcgcc
     4021 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     4081 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacacg tgctacgaga
     4141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4261 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4321 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4381 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat taccaatgct
     4441 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4501 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agcgctgcga
     4561 tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac cagccagccg
     4621 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4681 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4741 tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4801 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4861 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4921 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4981 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5041 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5101 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5161 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5221 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5281 gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5341 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa
     5401 ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac
     5461 accccttgct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     5521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5701 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     5761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6181 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6241 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6301 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     6361 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     6421 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     6481 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     6541 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     6601 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     6661 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     6721 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     6781 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     6841 cagtatttat tatcatttag aataaatttt gtgtcgccct taattgtgag cggataacaa
     6901 ttacgagctt catgcacagt ggcgttgact caatattggc cattagccat attattcatt
     6961 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     7021 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgtt
//
