LOCUS       sequence-127240-        5846 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 5846)
  AUTHORS   Mammoto A, Connor KM, Mammoto T, Yung CW, Huh D, Aderman CM, 
            Mostoslavsky G, Smith LE, Ingber DE
  TITLE     A mechanosensitive transcriptional mechanism that controls 
            angiogenesis.
  JOURNAL   Nature. 2009 Feb 26. 457(7233):1103-8.
  PUBMED    19242469
REFERENCE   2  (bases 1 to 5846)
  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: "Nature. 
            2009 Feb 26. 457(7233):1103-8."
COMMENT     Sequence Label: pGL3-VEGFR2-780
FEATURES             Location/Qualifiers
     source          1..5846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1933..2388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2020..2039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2230..2251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2519..2567
                     /note="synthetic polyadenylation signal"
     misc_feature    2581..2672
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2621..2640
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      3789..3798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3795..5447
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3861..3879)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5488..5609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5499..5518)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5553..5572
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat
     2401 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     2461 ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa
     2521 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     2581 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
     2641 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct ccctccttcc
     2701 cctgggccta aggatatctt ggctggaagc tctgctctga aaaggggcat ggccaaactt
     2761 tcactagggc tcttcgttgg ggagcacgat ggacaaaagc cttcttgggg ctaggcaggt
     2821 cacttcaaac ttggagccgc caaatatttt gggaaatagc gggaatgttg gcgaactggg
     2881 caagtgcgtt ttctgattaa gagcaaccag attcagcttt ttaaactaca attatactgg
     2941 ccaaacaaaa tacccttata caaaaaccaa aactactggc aggagtcgct gccagcttgc
     3001 gacccggcat acttggctga gtatccgctt ctcccttgtg gctccaaact gctgcagatt
     3061 ctcggccact tcagacgcgc gcgatggcga agagggtcct gcactttgac gcgcctggtg
     3121 agggagcgct gctcttcgca gcgctcctgg tgatgctccc caaatttcgg ggaccggcaa
     3181 gcgattaaat cttggagttg ctcagcgccc gttaccgagt actttttatt tacaccagaa
     3241 acaaagttgt tgctctggga tgttctctcc tgggcgactt ggggcccagc gcagtccagt
     3301 tgtgtgggga aatggggaga tgtaaatggg cttggggagc tggagatccc cgccgggtac
     3361 ccgggtgagg ggcggggctg gccgcacggg agagcccctc ctccgctccg gccccgcccc
     3421 gcatggcccc gcctccgcgc tctagagttt cggctccagc tcccaccctg cactgagtcc
     3481 cgggaccccg ggagagcggt cagtgtgtgg tcgctgcgtt tcctctgcct gcgccgggca
     3541 tcacttgcgc gccgcagaaa gtccgtctgg cagcctggat atcctctcct accggcaccc
     3601 gcagacgccc ctgcagccgc ggtcggcgcc cgggctccct agccctgtgc gctcaactgt
     3661 cctgcgctgc ggggtgccgc gagttccacc tccgcgcctc cttctctaga caggcgctgg
     3721 gagaaagaac cggctcccct cgagatctgc gatctaagta agcttggcat tccggtactg
     3781 ttggtaaagc caccatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
     3841 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
     3901 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
     3961 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     4021 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     4081 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     4141 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     4201 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
     4261 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     4321 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     4381 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     4441 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     4501 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     4561 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     4621 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     4681 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     4741 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     4801 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     4861 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     4921 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     4981 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     5041 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     5101 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     5161 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     5221 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     5281 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     5341 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     5401 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg
     5461 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     5521 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5581 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     5641 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     5701 aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     5761 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     5821 ctcgtaggac aggtgccggc agcgct
//
