LOCUS       sequence-10305-e        4660 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 4660)
  AUTHORS   Lawson ND, Vogel AM, Weinstein BM
  TITLE     sonic hedgehog and vascular endothelial growth factor act upstream 
            of the Notch pathway during arterial endothelial differentiation.
  JOURNAL   Dev Cell. 2002 Jul . 3(1):127-36.
  PUBMED    12110173
REFERENCE   2  (bases 1 to 4660)
  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: "Dev Cell. 
            2002 Jul . 3(1):127-36."
COMMENT     Sequence Label: pCS2vegf121
FEATURES             Location/Qualifiers
     source          1..4660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    709..843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(733..752)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     787..806
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(956..976)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(956..974)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(995..1011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(995..1011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1008..1030)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1019..1035
                     /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(1043..1049,1050..1067,1068..1073))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1088..1109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1226..1243)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1397..1985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1477..1496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2156..2947,2948..3016))
                     /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     2779..2798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3017..3121)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3147..3602)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3284..3305)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3496..3515
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3676..4660
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccacaatc atgaacttgg ttgtttattt gatacagtta
      121 tttctcgcgg ctctcctcca tctgtctgct gtaaaggctg cccacatacc caaagaaggg
      181 ggaaagagca aaaatgatgt gattcccttc atggatgtgt ataaaaagag tgcgtgcaag
      241 acccgagagc tgctggtaga catcatccag gagtatcccg atgagatcga gcacacgtac
      301 atcccgtcct gtgtggttct catgcgctgt gcaggatgct gtaatgatga ggcgctcgaa
      361 tgcgtcccga cagagacacg aaacgtcact atggaggtgc tgcgggtcaa gcaacgcgta
      421 tcgcagcata attttcagct gagtttcaca gaacacacca agtgtgaatg caggccaaag
      481 gcagaagtca aagcaaagaa agaaaaatgt gaaaagccaa gatgatgatc gagcgaacgc
      541 cgggcagaac tctgcatgaa ggaaggaact taaataaaaa ctttttaaac ggcacagcac
      601 tttcaaaacc taaggaagaa cattccttgg atgaactgca aacactatcc taaacgaaac
      661 actaagtgca gctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg
      721 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
      781 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
      841 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
      901 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
      961 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1021 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1081 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1141 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1201 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1261 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1321 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1801 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1921 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1981 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2101 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2161 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2221 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2281 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2341 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2401 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2461 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2521 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2581 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2641 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2701 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2761 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2821 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2881 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2941 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3001 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3061 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3121 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     3181 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3241 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3301 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3361 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3421 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3481 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3541 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3601 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3661 gctattacgc cagtcgacca tagccaattc aatatggcgt atatggactc atgccaattc
     3721 aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc
     3781 aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc
     3841 aatatggcgg acttggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt
     3901 ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata
     3961 atggcggcca tattggctat atgccaggat caatatatag gcaatatcca atatggccct
     4021 atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat
     4081 tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac
     4141 catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct
     4201 atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc
     4261 cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag
     4321 gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc
     4381 cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat
     4441 agtaacttgg caagtacatt actattggaa ggacgccagg gtacattggc agtactccca
     4501 ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca
     4561 atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc
     4621 ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
