LOCUS       sequence-71928-e        5259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression vector, In-situ probe template and in vitro 
            transcription vector for zebrafish Etv2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5259)
  AUTHORS   Moore JC, Sheppard S, Shestopalov IA, Chen JK, Lawson N
  TITLE     Post-transcriptional mechanisms contribute to Etv2 repression during
            vascular development.
  JOURNAL   Dev Biol. 2013 Sep 11. pii: S0012-1606(13)00470-3. doi: 
            10.1016/j.ydbio.2013.08.028.
  PUBMED    24036310
REFERENCE   2  (bases 1 to 5259)
  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 Biol. 
            2013 Sep 11. pii: S0012-1606(13)00470-3. doi: 
            10.1016/j.ydbio.2013.08.028."
COMMENT     Sequence Label: 645 pCS2-etv2
FEATURES             Location/Qualifiers
     source          1..5259
                     /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"
     protein_bind    108..132
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1239..1263)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1308..1442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1332..1351)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1386..1405
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1555..1575)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1555..1573)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1594..1610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1594..1610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1607..1629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1618..1634
                     /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(1642..1648,1649..1666,1667..1672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1687..1708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1825..1842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1996..2584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2076..2095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2755..3546,3547..3615))
                     /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     3378..3397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3616..3720)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3746..4201)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3883..3904)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4095..4114
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4275..5259
                     /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 atcccatcga ttcgaattca aggatcaaca agtttgtaca
      121 aaaaagcagg cttaaccatg gaaatgtacc aatctggatt ttacacagaa gacttcagaa
      181 ctcaggaggt tcctgctggt ttcgacttca gttcatatga ctgcagtggt gaagacctgt
      241 cctttttatt agacagcaaa ggacccgtcc aacagcagta tgccgaaaac tactccgagc
      301 ctcagaagga gctgttgcac aaaggtcatg ttctcacagt cgactctggg ctgttcaatc
      361 tggactcttt ccctgagttc agcaactggg cagcatacac taatattcca gaaggaatgg
      421 ttgcagacag acagcaggtt ggctttcagg aatcaactca aacctaccag aacctcgtcc
      481 ctctctgcac tccagcacaa agcagtacat tcagtcccac aatggacacc agcagccatt
      541 accaacctgg gaaaggccca agtcacagag gagcatccgg gaccgccagc cttgaccatt
      601 tgggtgaatc agacagaaca tatggtttgt atgaagcaga gcagcaaagc aggtcttcat
      661 attggtccga ctacccctca cccggttact gcagctccat gccgctgagc cagcctgcat
      721 cctcctcatc ccctccggtc agtcagtccg ctgagcactt ctgcccccgc gtggtcaaaa
      781 gacgcagcgc tcctcctcaa agatcagaca gagagggcga gatcacgcca atgtccgcct
      841 acccaggatc tggacccatc cagctttggc agtttctgct agaactcctg ctggattctg
      901 cttgccacac ttttataagt tggactggtg atggctggga gtttaaaatg tcagatcccg
      961 ctgaggtggc gaagcggtgg ggccagtgta aaaacaagcc taaaatgaac tatgagaagc
     1021 tcagtcgtgg cctgcgttac tactaccaca aaaacatcat tcacaaaacg gcgggaaagc
     1081 gctacgtcta ccgctttgtc tgtgacgtgc agggcatgct tggaaaaact gcacacgaag
     1141 tcttagcaag tctaaacatc tccccgaatg cagcatctcc acagtctgta gcaaacacaa
     1201 gccgctcaga ggaaaccaca gagtcctgga cacattagac ccagctttct tgtacaaagt
     1261 ggttgatcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
     1321 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1381 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1441 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1501 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     1561 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1621 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1681 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1741 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1801 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2401 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     2461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2521 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2641 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2701 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2761 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2821 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2881 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2941 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3001 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3061 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3121 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3181 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3241 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3301 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3361 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3421 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3481 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3541 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3601 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3661 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3721 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     3781 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3841 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3901 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3961 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4021 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4081 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4141 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4201 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4261 ctattacgcc agtcgaccat agccaattca atatggcgta tatggactca tgccaattca
     4321 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     4381 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     4441 atatggcgga cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     4501 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     4561 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     4621 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     4681 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     4741 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     4801 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     4861 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     4921 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     4981 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     5041 gtaacttggc aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat
     5101 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     5161 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     5221 taatgggagg tctatataag caatgctcgt ttagggaac
//
