LOCUS       sequence-369699-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of full length human tissue factor (CD142; F3) in a 
            Sleeping Beauty transposon vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  AUTHORS   Muhsin-Sharafaldine MR, Saunderson SC, Dunn AC, Faed JM, Kleffmann 
            T, McLellan AD
  TITLE     Procoagulant and immunogenic properties of melanoma exosomes, 
            microvesicles and apoptotic vesicles.
  JOURNAL   Oncotarget. 2016 Aug 30;7(35):56279-56294. doi: 
            10.18632/oncotarget.10783.
  PUBMED    27462921
REFERENCE   2  (bases 1 to 7284)
  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: 
            "10.18632/oncotarget"; journalName: "Oncotarget"; date: "2016-08-30-
            30"; volume: "7"; issue: "35"; pages: "56279-56294"
COMMENT     Sequence Label: pSBbiRP Hu Full Length CD142 (Tissue Factor; F3)
FEATURES             Location/Qualifiers
     source          1..7284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..60)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    49..273
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        705..809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(810..878,879..1670)
                     /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(1028..1047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1841..2429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2330..2349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2583..2600
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2974..2993
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2992..3047
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3093..3112
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(3212..3811)
                     /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(3295..3315)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3792..3811
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3812..3868)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(3878..4579)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3961..3984)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4159..4178)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4428..4446
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        5211..6393
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     6341..6361
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      6419..6428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     6425..6484
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
ORIGIN
        1 aggccaagct tggggatcaa ttctctagag ctcgctgatc agcctcgact gtgccttcta
       61 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
      121 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
      181 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
      241 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
      301 cctactagat agcggacccc ttaccgaaac atcgccgcat tctgcagagg agtcgaactt
      361 agtgtatgta aacttctgac ccactggaat tgtgatacag tgaattataa gtgaaataat
      421 ctgtctgtaa acaattgttg gaaaaatgac ttgtgtcatg cacaaagtag atgtcctaac
      481 tgacttgcca aaactattgt ttgttaacaa gaaatttgtg gagtagttga aaaacgagtt
      541 ttaatgactc caacttaagt gtatgtaaac ttccgacttc aactgtatag ggatccaatt
      601 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
      661 cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      721 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      781 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      841 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      901 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
      961 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1021 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1081 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1141 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1201 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1261 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1321 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1381 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1441 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1501 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1561 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1621 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1681 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2521 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2581 gcagcgagtc agtgagcgag gaagcggatc cctatacagt tgaagtcgga agtttacata
     2641 caccttagcc aaatacattt aaactcactt tttcacaatt cctgacattt aatcctagta
     2701 aaaattccct gtcttaggtc agttaggatc accactttat tttaagaatg tgaaatatca
     2761 gaataatagt agagagaatg attcatttca gcttttattt ctttcatcac attcccagtg
     2821 ggtcagaagt ttacatacac tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg
     2881 tcccgagtag ctagttcatg gcagccagca tatggcatat gttgccaaac tctaaaccaa
     2941 atactcattc tgatgtttta aatgatttgc cctcccatat gtccttccga gtgagagaca
     3001 caaaaaattc caacacacta ttgcaatgaa aataaatttc ctttattagc cagaagtcag
     3061 atgctcaagg ggcttcatga tgtccccata atttttggca gagggaaaaa gatctcagtg
     3121 gtatttgtga gccagggcat tggccacacc agccaccacc ttctgatagg cagcctgcac
     3181 ctgaggagtg aattcacgac aggccttcga atcaggcacc gggcttgcgg gtcatgcacc
     3241 aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt
     3301 agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg
     3361 ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg
     3421 agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc
     3481 cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga
     3541 tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg
     3601 cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt
     3661 cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg
     3721 ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg
     3781 cgaggcgcac cgtgggcttg tactcggtca tggggccagg attctcctcg acgtcaccag
     3841 cctgcttcag caggctgaag ttagtagctc cactgccctt gtacagctcg tccatgccgt
     3901 acaggaacag gtggtggcgg ccctcggagc gctcgtactg ttccacgatg gtgtagtcct
     3961 cgttgtggga ggtgatgtcc agcttggtgt ccacgtagta gtagccgggc agttgcacgg
     4021 gcttcttggc catgtagatg gtcttgaact ccaccaggta gtggccgccg tccttcagct
     4081 tcagggcctg gtggatctcg cccttcagca cgccgtcgcg ggggtacagg cgctcggtgg
     4141 aggcctccca gcccattgtc ttcttctgca ttacggggcc gtcggggggg aagttggtgc
     4201 cgcgcatctt caccttgtag atcagcgtgc cgtcctgcag ggaggagtcc tgggtcacgg
     4261 tcaccagacc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
     4321 acagcttctt gtaatcgggg atgtcggcgg ggtgcttcac gtacgccttg gagccgtaca
     4381 tgaactgggg ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca
     4441 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
     4501 catggccgtt catggagccc tccatgcgca ccttgaagcg catgaactct ttgatgacct
     4561 cctcgccctt gctcaacatg gtggcctcag gtgcagaggt ttctacaggg aaagagagaa
     4621 aggctacagt tagtttgctg tgatgatgct aagtgccgag gtcagatgtc tatggattgt
     4681 acacggaatc ccaacgcagg aaccaaaaaa cactcaccca aaaaaatggc gctaaggccg
     4741 agagaaaggc gaagttccgt ctacggctat ataccttgaa acaggggcgg cagaagtgac
     4801 gcaacgcggt tgcgcggacg gcttcctgtg ttttcattgg tcgaagcgtt ctgcgcgtgc
     4861 gcagagattc cccagggaag gcgggccctc ttccgaatgt ccaatccgga cattcttccg
     4921 gttggacctc cgcggtcagc gctgtgcccg gcgagagatc acgtggggcg cggaggcggt
     4981 gctgctgggg cacggccgtc cagcctcggc ggccatattt ttgaggggct gttcatctcg
     5041 ttcacacgct ctgtccgcca tgtttgtgag tggaagcgcc attacccctt caagcgactg
     5101 aaggctgcag ggcctctggt ggcccgcatg gggagaccag acccgccagg cccgcctttc
     5161 cgcactcagt ccgggcttac tttattttgt gagacagggt ctcgcctaga ggctccggtg
     5221 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     5281 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     5341 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     5401 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
     5461 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacgc
     5521 ccctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
     5581 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcttggg
     5641 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     5701 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     5761 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     5821 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     5881 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     5941 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     6001 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     6061 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     6121 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     6181 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     6241 ccccatactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     6301 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     6361 caaagttttt ttcttccatt taaggtgtcg tgaaaactac cccaagctgg cctctgaggc
     6421 caccatggag acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac
     6481 tggttcaggc actacaaata ctgtggcagc atataattta acttggaaat caactaattt
     6541 caagacaatt ttggagtggg aacccaaacc cgtcaatcaa gtctacactg ttcaaataag
     6601 cactaagtca ggagattgga aaagcaaatg cttttacaca acagacacag agtgtgacct
     6661 caccgacgag attgtgaagg atgtgaagca gacgtacttg gcacgggtct tctcctaccc
     6721 ggcagggaat gtggagagca ccggttctgc tggggagcct ctgtatgaga actccccaga
     6781 gttcacacct tacctggaga caaacctcgg acagccaaca attcagagtt ttgaacaggt
     6841 gggaacaaaa gtgaatgtga ccgtagaaga tgaacggact ttagtcagaa ggaacaacac
     6901 tttcctaagc ctccgggatg tttttggcaa ggacttaatt tatacacttt attattggaa
     6961 atcttcaagt tcaggaaaga aaacagccaa aacaaacact aatgagtttt tgattgatgt
     7021 ggataaagga gaaaactact gtttcagtgt tcaagcagtg attccctccc gaacagttaa
     7081 ccggaagagt acagacagcc cggtagagtg tatgggccag gagaaagggg aattcagaga
     7141 aatattctac atcattggag ctgtggcatt tgtggtcatc atccttgtca tcatcctggc
     7201 tatatctcta cacaagtgta gaaaggcagg agtggggcag agctggaagg agaactcccc
     7261 actgaatgtt tcataaggcc tgtc
//
