LOCUS       sequence-125131-        6698 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 6698)
  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 6698)
  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: -948 hTF
FEATURES             Location/Qualifiers
     source          1..6698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1177..2829
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1243..1261)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          3069..3134
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3264..3284
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3709..3843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3985..4002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4156..4744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4236..4255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4915..5706,5707..5775))
                     /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     5538..5557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5776..5880)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5907..6362
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5994..6013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6204..6225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6552..6673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cccagcttgc atgcctgcag gtcgactcta gaggatcccc agcttaaaac tagtggcaca
       61 caactctggt tacatttttc attttcattt ttggttcctc tgcatttcgg tggaactcag
      121 tcccacagat gtcttcttca atgtgggaac catcctttct ggaaccacag agctgcagat
      181 gtcacgctgg aattctccca gaggcaaact gccagatgtg aggctgctct tcctcagtca
      241 ctatctctgg tcgtaccggg cgatgcctga gccaactgac cctcagacct gtgagccgag
      301 ccggtcacac cgtggctgac accggcattc ccaccgcctt tctcctgtgc gacccgctaa
      361 gggccccgcg aggtgggcag gccaagtatt cttgaccttc gtggggtaga agaagccacc
      421 gtggctggga gagggccctg ctcacagcca cacgtttact tcgctgcagg tcccgagctt
      481 ctgccccagg tgggcaaagc atccgggaaa tgccctccgc tgcccgaggg gagcccagag
      541 cccgtgcttt ctattaaatg ttgtaaatgc cgcctctccc actttatcac caaatggaag
      601 ggaagaattc ttccaaggcg ccctcccttt cctgccatag acctgcaacc cacctaagct
      661 gcacgtcgga gtcgcgggcc tgggtgaatc cgggggcctt gggggacccg ggcaactaga
      721 cccgcctgcg tcctccaggg cagctccgcg ctcggtggcg cggttgaatc actggggtga
      781 gtcatccctt gcagggtccc ggagtttcct accgggagga ggcggggcag gggtgtggac
      841 tcgccggggg ccgcccaccg cgacggcaag tgacccgggc cgggggcggg gagtcgggag
      901 gagcggcggg ggcgggcgcc gggggcgggc agaggcgcgg gagagcgcgc cgccggccct
      961 ttatagcgcg cggggcaccg gctccccaag actgcgagct ccccgcaccc cctcgcactc
     1021 cctctggccg gcccagggcg ccttcagccc aacctcccca gccccacggg cgccacggaa
     1081 cccgctcgat ctcgccgcca actggtaggg taccgagctc ttacgcgtgc tagctcgaga
     1141 tctaagtaag cttggcattc cggtactgtt ggtaaaatgg aagacgccaa aaacataaag
     1201 aaaggcccgg cgccattcta tcctctagag gatggaaccg ctggagagca actgcataag
     1261 gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag
     1321 gtgaacatca cgtacgcgga atacttcgaa atgtccgttc ggttggcaga agctatgaaa
     1381 cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc
     1441 tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt
     1501 tataatgaac gtgaattgct caacagtatg aacatttcgc agcctaccgt agtgtttgtt
     1561 tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa aattaccaat aatccagaaa
     1621 attattatca tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc
     1681 acatctcatc tacctcccgg ttttaatgaa tacgattttg taccagagtc ctttgatcgt
     1741 gacaaaacaa ttgcactgat aatgaattcc tctggatcta ctgggttacc taagggtgtg
     1801 gcccttccgc atagaactgc ctgcgtcaga ttctcgcatg ccagagatcc tatttttggc
     1861 aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga
     1921 atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt
     1981 gaagaagagc tgtttttacg atcccttcag gattacaaaa ttcaaagtgc gttgctagta
     2041 ccaaccctat tttcattctt cgccaaaagc actctgattg acaaatacga tttatctaat
     2101 ttacacgaaa ttgcttctgg gggcgcacct ctttcgaaag aagtcgggga agcggttgca
     2161 aaacgcttcc atcttccagg gatacgacaa ggatatgggc tcactgagac tacatcagct
     2221 attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt
     2281 tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcagagaggc
     2341 gaattatgtg tcagaggacc tatgattatg tccggttatg taaacaatcc ggaagcgacc
     2401 aacgccttga ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa
     2461 gacgaacact tcttcatagt tgaccgcttg aagtctttaa ttaaatacaa aggatatcag
     2521 gtggcccccg ctgaattgga atcgatattg ttacaacacc ccaacatctt cgacgcgggc
     2581 gtggcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag
     2641 cacggaaaga cgatgacgga aaaagagatc gtggattacg tggccagtca agtaacaacc
     2701 gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga
     2761 aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagtcc
     2821 aaattgtaaa atgtaactgt attcagcgat gacgaaattc ttagctattg taatactgcg
     2881 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     2941 gtgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcgc cggatctttg
     3001 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
     3061 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg
     3121 tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct
     3181 ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg
     3241 ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact
     3301 ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt
     3361 gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa
     3421 aatattctgt aacctttata agtaggcata acagttataa tcataacata ctgttttttc
     3481 ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct
     3541 ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta
     3601 gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3661 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3721 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3781 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     3841 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3901 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3961 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4321 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     4381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4561 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4981 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5041 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5101 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5161 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5221 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5281 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5341 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5401 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5461 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5521 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5581 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5641 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5701 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5761 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5821 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5881 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5941 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6001 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6061 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6121 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6181 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6241 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6301 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     6361 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6421 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gtggaggttt
     6481 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     6541 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6601 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6661 tcaatgtatc ttatggtact gtaactgagc taacataa
//
