LOCUS       sequence-191798-        5788 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 5788)
  AUTHORS   Carson-Walter EB, Watkins DN, Nanda A, Vogelstein B, Kinzler KW, St 
            Croix B
  TITLE     Cell surface tumor endothelial markers are conserved in mice and 
            humans.
  JOURNAL   Cancer Res. 2001 Sep 15. 61(18):6649-55.
  PUBMED    11559528
REFERENCE   2  (bases 1 to 5788)
  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: "Cancer Res.
            2001 Sep 15. 61(18):6649-55."
COMMENT     Sequence Label: pIVEX2.3 TEM-5
FEATURES             Location/Qualifiers
     source          1..5788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2207..2224
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2318..2336)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2332..2379
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2745..2750,2751..2767,2768..2773))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2794..2810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2794..2810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2807..2829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2818..2834
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2842..2848,2849..2866,2867..2872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2887..2908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3025..3042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3196..3784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3276..3295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3955..4746,4747..4815))
                     /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     4578..4597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4816..4920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4988..5006
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5044..5066)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5166..5185
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5381..5403
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5395..5412
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5396..5412
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5532..5551
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5635..5654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5635..5653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             5685..5707
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 gctcctggcg cccgaggctc ggggcgcgcc cggctgcccg ctatccatcc gcagctgcaa
       61 gtgctcgggg gagcggccca aggggctgag cggcggcgtc cctggcccgg ctcggcggag
      121 ggtggtgtgc agcggcgggg acctcccgga gcctcccgag cccggccttc tgcctaacgg
      181 caccgttacc ctgctcttga gcaataacaa gatcacgggg ctccgcaatg gctccttcct
      241 gggactgtca ctgctggaga agctggacct gaggaacaac atcatcagca cagtgcagcc
      301 gggcgccttc ctgggcctgg gggagctgaa gcgtttagat ctctccaaca accggattgg
      361 ctgtctcacc tccgagacct tccagggcct ccccaggctt ctccgactaa acatatctgg
      421 aaacatcttc tccagtctgc aacctggggt ctttgatgag ctgccagccc ttaaggttgt
      481 ggacttgggc accgagttcc tgacctgtga ctgccacctg cgctggctgc tgccctgggc
      541 ccagaatcgc tccctgcagc tgtcggaaca cacgctctgt gcttacccca gtgccctgca
      601 tgctcaggcc ctgggcagcc tccaggaggc ccagctctgc tgcgaggggg ccctggagct
      661 gcacacacac cacctcatcc cgtccctacg ccaagtggtg ttccaggggg atcggctgcc
      721 cttccagtgc tctgccagct acctgggcaa cgacacccgc atccgctggt accacaaccg
      781 agcccctgtg gagggtgatg agcaggcggg catcctcctg gccgagagcc tcatccacga
      841 ctgcaccttc atcaccagtg agctgacgct gtctcacatc ggcgtgtggg cctcaggcga
      901 gtgggagtgc accgtgtcca tggcccaagg caacgccagc aagaaggtgg agatcgtggt
      961 gctggagacc tctgcctcct actgccccgc cgagcgtgtt gccaacaacc gcggggactt
     1021 caggtggccc cgaactctgg ctggcatcac agcctaccag tcctgcctgc agtatccctt
     1081 cacctcagtg cccctgggcg ggggtgcccc gggcacccga gcctcccgcc ggtgtgaccg
     1141 tgccggccgc tgggagccag gggactactc ccactgtctc tacaccaacg acatcaccag
     1201 ggtgctgtac accttcgtgc tgatgcccat caatgcctcc aatgcgctga ccctggctca
     1261 ccagctgcgc gtgtacacag ccgaggccgc tagcttttca gacatgatgg atgtagtcta
     1321 tgtggctcag atgatccaga aatttttggg ttatgtcgac cagatcaaag agctggtaga
     1381 ggtgatggtg gacatggcca gcaacctgat gctggtggac gagcacctgc tgtggctggc
     1441 ccagcgcgag gacaaggcct gcagccgcat cgtgggtgcc ctggagcgca ttgggggggc
     1501 cgccctcagc ccccatgccc agcacatctc agtgaatgcg aggaacgtgg cattggaggc
     1561 ctacctcatc aagccgcaca gctacgtggg cctgacctgc acagccttcc agaggaggga
     1621 gggaggggtg ccgggcacac ggccaggaag ccctggccag aaccccccac ctgagcccga
     1681 gcccccagct gaccagcagc tccgcttccg ctgcaccacc gggaggccca atgtttctct
     1741 gtcgtccttc cacatcaaga acagcgtggc cctggcctcc atccagctgc ccccgagtct
     1801 attctcatcc cttccggctg ccctggctcc cccggtgccc ccagactgca ccctgcaact
     1861 gctcgtcttc cgaaatggcc gcctcttcca cagccacagc aacacctccc gccctggagc
     1921 tgctgggcct ggcaagaggc gtggcgtggc cacccccgtc atcttcgcag gaaccagtgg
     1981 ctgtggcgtg ggaaacctga cagagccagt ggccgtttcg ctgcggcact gggctgaggg
     2041 agccgaacct gtggccgctt ggtggagcca ggaggggccc ggggaggctg ggggctggac
     2101 ctcggagggc tgccagctcc gctccagcca gcccaatgtc agcgccctgc actgccagca
     2161 cttgggcaat gtggccgtgc tcatggagct ggatgggggg ggttctcatc atcatcatca
     2221 tcattaataa aagggcgaat tccagcacac tggcggccgt tactagtgga tccggctgct
     2281 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2341 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     2401 ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag
     2461 cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc
     2521 gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
     2581 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     2641 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     2701 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     2761 gataagctgt caaacatgag aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     2821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3001 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3061 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4921 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4981 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5041 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5101 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5161 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg
     5221 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     5281 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     5341 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     5401 acgacggcca gtgccaagct tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     5461 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     5521 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     5581 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     5641 gactcactat agggagacca caacggtttc cctctagaaa taattttgtt taactttaag
     5701 aaggagatat acatatgggc gccgggggac gcaggatgcg gggggcgccc gcgcgcctgc
     5761 tgctgccgct gctgccgtgg ctcctgct
//
