LOCUS       sequence-369717-        6849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Soluble expression of the extracellular domains of human tissue 
            factor (TF)-Fc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6849)
  TITLE     McLellan Lab Plasmids 2022
REFERENCE   2  (bases 1 to 6849)
  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"
COMMENT     Sequence Label: pcDNA3.1(-) HuTF-Fc-His tag
FEATURES             Location/Qualifiers
     source          1..6849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..43)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    32..256
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      302..730
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(389..408)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     599..620
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(739..759)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        744..1073
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      924..1059
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     986..1005
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1140..1934
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1194..1213)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1804..1823
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2108..2229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2145..2164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2199..2218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2278..2294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2278..2294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2291..2313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2302..2318
                     /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)."
     promoter        complement(join(2326..2332,2333..2350,2351..2356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2371..2392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2509..2526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2680..3268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2760..2779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3439..4230,4231..4299))
                     /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     4062..4081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4300..4404)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4479..4498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4670..5049
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5050..5253
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5204..5224
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5298..5317
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5298..5316
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6809..6832
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
ORIGIN
        1 agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
       61 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
      121 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
      181 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
      241 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
      301 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      361 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      421 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      481 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      541 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      601 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      661 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      721 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      841 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      901 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      961 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     1021 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     1081 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     1141 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     1201 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     1261 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     1321 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     1381 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     1441 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     1501 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     1561 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     1621 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     1681 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     1741 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     1801 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     1861 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     1921 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     1981 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     2041 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     2101 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2161 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2221 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     2281 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2341 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2401 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     4441 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     4501 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     4561 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     4621 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     4681 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     4741 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4801 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     4861 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     4921 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4981 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     5041 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     5101 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     5161 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     5221 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     5281 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
     5341 aacgggccct ctagactcga gcggccgcca ctgtgctgga tatctgcaga attcggcctc
     5401 tgaggccacc atgcgaagga tgcaacttct tctgctcatc gcactttcct tggcgctggt
     5461 gacaaatagt actacaaata ctgtggcagc atataattta acttggaaat caactaattt
     5521 caagacaatt ttggagtggg aacccaaacc cgtcaatcaa gtctacactg ttcaaataag
     5581 cactaagtca ggagattgga aaagcaaatg cttttacaca acagacacag agtgtgacct
     5641 caccgacgag attgtgaagg atgtgaagca gacgtacttg gcacgggtct tctcctaccc
     5701 ggcagggaat gtggagagca ccggttctgc tggggagcct ctgtatgaga actccccaga
     5761 gttcacacct tacctggaga caaacctcgg acagccaaca attcagagtt ttgaacaggt
     5821 gggaacaaaa gtgaatgtga ccgtagaaga tgaacggact ttagtcagaa ggaacaacac
     5881 tttcctaagc ctccgggatg tttttggcaa ggacttaatt tatacacttt attattggaa
     5941 atcttcaagt tcaggaaaga aaacagccaa aacaaacact aatgagtttt tgattgatgt
     6001 ggataaagga gaaaactact gtttcagtgt tcaagcagtg attccctccc gaacagttaa
     6061 ccggaagagt acagacagcc cggtagagtg tatgggccag gagaaagggg agttcagaga
     6121 aggtaccgac aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc
     6181 gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga
     6241 ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta
     6301 cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag
     6361 cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
     6421 gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa
     6481 agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat
     6541 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
     6601 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
     6661 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
     6721 gcaggggaac gtcttctcat gctccgtgat gcacgaggct ctgcacaacc actacacgca
     6781 gaagagcctc tccctgtctc cgggtaaaca tcaccaccat caccatcatc actgaggcct
     6841 gtcaggcca
//
