LOCUS       sequence-125136-        5877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5877)
  AUTHORS   Mackman N
  TITLE     Regulation of the tissue factor gene.
  JOURNAL   FASEB J. 1995 Jul . 9(10):883-9.
  PUBMED    7615158
REFERENCE   2  (bases 1 to 5877)
  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"; journalName: "FASEB J. 
            1995 Jul . 9(10):883-9."
COMMENT     Sequence Label: -278 hTF
FEATURES             Location/Qualifiers
     source          1..5877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             356..2008
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(422..440)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2248..2313
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2443..2463
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2888..3022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3164..3181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3335..3923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3415..3434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4094..4885,4886..4954))
                     /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     4717..4736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4955..5059)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5086..5541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5173..5192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5383..5404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5731..5852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cccgactcta gagggcaact agacccgcct gcgtcctcca gggcagctcc gcgctcggtg
       61 gcgcggttga atcactgggg tgagtcatcc cttgcagggt cccggagttt cctaccggga
      121 ggaggcgggg caggggtgtg gactcgccgg gggccgccca ccgcgacggc aagtgacccg
      181 ggggcgggga gtcgggagga gcggcggggg cgggcgccgg gggcgggcag aggcgcggga
      241 gagcgcgccg ccggcccttt atagcgcgcg gggcaccggc tccccaagac tgcgagctct
      301 tacgcgtgct agctcgagat ctaagtaagc ttggcattcc ggtactgttg gtaaaatgga
      361 agacgccaaa aacataaaga aaggcccggc gccattctat cctctagagg atggaaccgc
      421 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
      481 tacagatgca catatcgagg tgaacatcac gtacgcggaa tacttcgaaa tgtccgttcg
      541 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
      601 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
      661 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatga acatttcgca
      721 gcctaccgta gtgtttgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
      781 attaccaata atccagaaaa ttattatcat ggattctaaa acggattacc agggatttca
      841 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
      901 accagagtcc tttgatcgtg acaaaacaat tgcactgata atgaattcct ctggatctac
      961 tgggttacct aagggtgtgg cccttccgca tagaactgcc tgcgtcagat tctcgcatgc
     1021 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     1081 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     1141 cgtcttaatg tatagatttg aagaagagct gtttttacga tcccttcagg attacaaaat
     1201 tcaaagtgcg ttgctagtac caaccctatt ttcattcttc gccaaaagca ctctgattga
     1261 caaatacgat ttatctaatt tacacgaaat tgcttctggg ggcgcacctc tttcgaaaga
     1321 agtcggggaa gcggttgcaa aacgcttcca tcttccaggg atacgacaag gatatgggct
     1381 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     1441 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     1501 gggcgttaat cagagaggcg aattatgtgt cagaggacct atgattatgt ccggttatgt
     1561 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     1621 catagcttac tgggacgaag acgaacactt cttcatagtt gaccgcttga agtctttaat
     1681 taaatacaaa ggatatcagg tggcccccgc tgaattggaa tcgatattgt tacaacaccc
     1741 caacatcttc gacgcgggcg tggcaggtct tcccgacgat gacgccggtg aacttcccgc
     1801 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     1861 ggccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     1921 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     1981 caagaagggc ggaaagtcca aattgtaaaa tgtaactgta ttcagcgatg acgaaattct
     2041 tagctattgt aatactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     2101 tggtgccctt aaacgcctgg tgctacgcct gaataagtga taataagcgg atgaatggca
     2161 gaaattcgcc ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa
     2221 ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt
     2281 aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg
     2341 gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct
     2401 agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag
     2461 gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt
     2521 agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta
     2581 tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat
     2641 cataacatac tgttttttct tactccacac aggcatagag tgtctgctat taataactat
     2701 gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg
     2761 atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag aggttttact
     2821 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2881 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2941 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3001 tgtatcttat catgtctgga tccgtcgacc gatgcccttg agagccttca acccagtcag
     3061 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     3121 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
     3181 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3241 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3301 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3361 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     3541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3721 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3781 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3841 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3901 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3961 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4021 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4081 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4141 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4201 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4261 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4321 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4381 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4441 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4501 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4561 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4621 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4681 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4741 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4801 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4861 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4921 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4981 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5041 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     5101 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5161 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5221 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5281 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5341 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5401 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5461 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5521 aaatattaac gcttacaatt tgccattcgc cattcaggct gcgcaactgt tgggaagggc
     5581 gatcggtgcg ggcctcttcg ctattacgcc agcccaagct accatgataa gtaagtaata
     5641 ttaaggtacg tggaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     5701 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5821 tgtggtttgt ccaaactcat caatgtatct tatggtactg taactgagct aacataa
//
