LOCUS       40924_44519        6572 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTVDll42AEmerald, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  AUTHORS   Blanco R, Geudens I, Stanchi F, Mathivet T, Jones M, Bentley K, 
            Gerhardt H.
  TITLE     Collective endothelial Dll4/Notch dynamics switch between vessel 
            branching and expansion
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6572)
  AUTHORS   Ubezio B.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2013) Vascular Biology Laboratory, Cancer Research
            UK, 44 Lincoln's Inn Fields, London, London WC2A 3LY, United Kingdom
REFERENCE   3  (bases 1 to 6572)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6572)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUN-2013) Vascular Biology Laboratory, Cancer Research UK, 44 
            Lincoln's Inn Fields, London, London WC2A 3LY, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        471..575
                     /label=AmpR promoter
     CDS             576..1433
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1607..2195
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2483..2504
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2519..2549
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2557..2573
                     /label=lac operator
                     /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)."
     primer_bind     2581..2597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2618..2636
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3215..3928
                     /codon_start=1
                     /label=EmGFP
                     /note="Emerald GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     protein_bind    4006..4039
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4104..4603
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        4615..4662
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4681..5481
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5522..5746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(6379..6397)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6404..6420)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(join(6561..6572,1..444))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
       61 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
      121 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
      181 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      241 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      301 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      361 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      421 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      481 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      541 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      661 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      841 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      961 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1081 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1141 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1261 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1441 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2281 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2341 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2401 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2461 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     2521 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     2581 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
     2641 aaaagctggt accgtgagtt cttccctcta agcatgtccc ctcctgcttt gtgtggtggg
     2701 aaaaaaatgt cctgttcact agcaatctca ttcctgaagg ggtgggtcag agatctcttc
     2761 tttggtgtgt agtggctctt gggtgccctg ctggccccat tgccacagag ggagtgatac
     2821 tggagccagg tggtatgcct gacttggcag ctgtgccagg gaaagggaca tagtcaagga
     2881 gcaggagaag gctcaggcag agctttcagg aactatttcc cacttgcctt tgggtgaaga
     2941 cacagggtta cctgtgtctg cttcctccat tgaactcctc ttgtgtgtct tagagcagga
     3001 ggctgagggt actaactccc tcagtggtgt ctcctagaga ggtccagagc acctctggat
     3061 cattgcatac cgcccccccc cacccccggt ataaccattt tcccattttg tatgtgatcc
     3121 ccaggtatta attaacgtcg agggcagcgg cgcgaccaac tttagcctgc tgaaacaggc
     3181 gggcgatgtg gaagaaaacc caggaccgac tagtatggtg agcaagggcg aggagctgtt
     3241 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3301 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3361 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt
     3421 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3481 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3541 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3601 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3661 caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg
     3721 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3781 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     3841 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3901 gatcactctc ggcatggacg agctgtactg agcggccgcg tcgacctgca gccaagctat
     3961 cgaattcctg cagcccaatt ccgatcatat tcaataaccc ttaatataac ttcgtataat
     4021 gtatgctata cgaagttatt aggtctgaag aggagtttac gtccagccaa gctagcttgg
     4081 ctgcaggtcg tcgaaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc
     4141 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
     4201 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
     4261 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
     4321 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     4381 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
     4441 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     4501 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
     4561 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcctgttgac
     4621 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     4681 atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4741 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4801 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4861 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4921 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4981 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5041 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5101 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5161 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5221 atgcccgacg gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5281 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5341 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5401 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5461 cgccttcttg acgagttctt ctgaggggat caattctcta gagctcgctg atcagcctcg
     5521 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5581 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5641 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5701 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     5761 agaaccagct ggggctcgac tagagcttgc ggaaccctta atataacttc gtataatgta
     5821 tgctatacga agttattagg tccctcgagg ggatccggca ggagcctact cagacaccca
     5881 gctccggccc agcagctggg ccttccttct gcattgttta cattgcatcc tgtatgggac
     5941 atctttagta tgcacagtgc tgctctgcgg aggaggagga aatggcatga actgaacaga
     6001 ctgtgaaccc gccaagagtc gcaccggctc tgcacacctc caggagtctg cctggcttca
     6061 gatgggcagc cccgccaagg gaacagagtt gaggagttag aggagcatca gttgagctga
     6121 tatctaaggt gcctctcgaa cttggacttg ctctgccaac agtggtcatc atggagctct
     6181 tgactgttct ccagagagtg gcagtggccc tagtgggtct tggcgctgct gtagctcctg
     6241 tgggcatctg tatttccaaa gtgcctttgc ccagactcca tcctcacagc tgggcccaaa
     6301 tgagaaagca gagaggaggc ttgcaaagga taggcctccc gcaggcagaa cagcctccgc
     6361 ggtggagctc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     6421 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6481 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6541 gcagcctgaa tggcgaatgg aaattgtaag cg
//
