LOCUS       sequence-172548-        5679 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 5679)
  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 5679)
  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: -67 hTF
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    151..272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             455..2107
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(521..539)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2348..2413
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2543..2563
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2988..3122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3264..3281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3435..4023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3515..3534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4194..4985,4986..5054))
                     /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     4817..4836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5055..5159)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5186..5641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5273..5292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5483..5504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat
       61 attaaggtac gtggaggttt tacttgcttt aaaaaacctc cacacctccc cctgaacctg
      121 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
      181 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
      241 tgtggtttgt ccaaactcat caatgtatct tatggtactg taactgagct aacataaccc
      301 gactctagag gatccccggg ggcgggcaga ggcgcgggag agcgcgccgc cggcccttta
      361 tagcgcgcgg ggcaccggct ccccaagact gcgagctctt acgcgtgcta gctcgagatc
      421 taagtaagct tggcattccg gtactgttgg taaaatggaa gacgccaaaa acataaagaa
      481 aggcccggcg ccattctatc ctctagagga tggaaccgct ggagagcaac tgcataaggc
      541 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
      601 gaacatcacg tacgcggaat acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
      661 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
      721 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
      781 taatgaacgt gaattgctca acagtatgaa catttcgcag cctaccgtag tgtttgtttc
      841 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaaa ttaccaataa tccagaaaat
      901 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
      961 atctcatcta cctcccggtt ttaatgaata cgattttgta ccagagtcct ttgatcgtga
     1021 caaaacaatt gcactgataa tgaattcctc tggatctact gggttaccta agggtgtggc
     1081 ccttccgcat agaactgcct gcgtcagatt ctcgcatgcc agagatccta tttttggcaa
     1141 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     1201 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     1261 agaagagctg tttttacgat cccttcagga ttacaaaatt caaagtgcgt tgctagtacc
     1321 aaccctattt tcattcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     1381 acacgaaatt gcttctgggg gcgcacctct ttcgaaagaa gtcggggaag cggttgcaaa
     1441 acgcttccat cttccaggga tacgacaagg atatgggctc actgagacta catcagctat
     1501 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     1561 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc agagaggcga
     1621 attatgtgtc agaggaccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     1681 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     1741 cgaacacttc ttcatagttg accgcttgaa gtctttaatt aaatacaaag gatatcaggt
     1801 ggcccccgct gaattggaat cgatattgtt acaacacccc aacatcttcg acgcgggcgt
     1861 ggcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     1921 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     1981 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     2041 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagtccaa
     2101 attgtaaaat gtaactgtat tcagcgatga cgaaattctt agctattgta atactgcgat
     2161 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
     2221 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgcc ggatctttgt
     2281 gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag
     2341 ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt
     2401 ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt
     2461 taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc
     2521 tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt
     2581 tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg
     2641 ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa
     2701 atattctgta acctttataa gtaggcataa cagttataat cataacatac tgttttttct
     2761 tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt
     2821 tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag
     2881 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2941 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3001 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3061 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3121 tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     3181 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     3241 tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3301 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3361 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3421 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3481 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3541 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3601 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3661 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3721 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3781 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3841 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3901 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3961 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4021 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4081 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4141 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4201 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4261 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4321 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4381 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4441 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4501 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4561 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4621 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4681 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4741 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4801 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4861 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4921 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4981 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5041 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5101 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5161 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5221 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5281 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5341 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5401 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5461 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5521 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5581 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5641 tgccattcgc cattcaggct gcgcaactgt tgggaaggg
//
