LOCUS       sequence-80477-e        3728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  actinodin 3 and3 antisense in situ probe linearize w/ ApaI SP6 pol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3728)
  AUTHORS   Nolte H, Holper S, Housley MP, Islam S, Piller T, Konzer A, Stainier
            DY, Braun T, Kruger M
  TITLE     Dynamics of zebrafish fin regeneration using a pulsed SILAC 
            approach.
  JOURNAL   Proteomics. 2015 Feb;15(4):739-51. doi: 10.1002/pmic.201400316.
  PUBMED    25504979
REFERENCE   2  (bases 1 to 3728)
  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"; doi: "10.1002/pmic"; 
            journalName: "Proteomics"; date: "2015-02"; volume: "15"; issue: 
            "4"; pages: "739-51"
COMMENT     Sequence Label: pGEM-and3 in situ probe
FEATURES             Location/Qualifiers
     source          1..3728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      33..42
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      37..46
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(853..871)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(854..871)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(889..905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(889..905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(902..924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    913..929
                     /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(937..943,944..961,962..967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    982..1003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1120..1137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1291..1879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1371..1390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2050..2841,2842..2910))
                     /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     2673..2692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2911..3015)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3093..3548)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3230..3251)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3442..3461
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3674..3696
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3688..3705
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3689..3705
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3712..3728,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3712..3728,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tcgtcagagg agatctgtgg atcccaactg gtatcgcaaa aatcccgact ttcagtccta
      121 ctatcgttac tataatagta ttgggcacac agaagggctc tatgagatcg ataaaattag
      181 aatgctttac cagcaaatgc gtcatctgga gctcacctat ggccctgacg catcgaaata
      241 ccagaatacc cttggtctaa agccgacccc tccacccaca actcagcctc caccccctac
      301 tttaccacct gtctccaaga tgaatgttat atacctgtgc aattcaaaag acccccagtg
      361 caagccacat attgtctacc tgcccgccgg tggtgtgcct gtactttgcg atccacgtta
      421 ccatccaaac tgtaaactaa gtcctgccca gaattctccc tcaccacctg taattgtccc
      481 agaaccagtg acgactgtcc ctcttccacc aacacaaaaa tccactccac caccacctcc
      541 tcctcctata atggttgtca aaggaatgga gtatgactgt gatccttact gggaccctga
      601 ctgcctgatc gataatcctc caaggccagt aaaaatggtt gaaccgtcat cagagctccc
      661 agatgagaag gtgaaaaagg aaattccagt cgatccagtg ccaacctatg acccatatga
      721 cttcaatcaa gacctgtatg acccctttcg ccatgctgct ccagctcact aaaatcacta
      781 gtgaattcgc ggccgcctgc aggtcgacca tatgggagag ctcccaacgc gttggatgca
      841 tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt catagctgtt
      901 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      961 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1021 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1081 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1141 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1201 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1261 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1321 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1381 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1441 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1501 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1561 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1621 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1681 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1741 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1801 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1861 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1921 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1981 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2041 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2101 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2161 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2221 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2281 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2341 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2401 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2461 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2521 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2581 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2641 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2701 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2761 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2821 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2881 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2941 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3001 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt gaaataccgc
     3061 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     3121 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3181 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3241 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3301 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3361 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3421 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3481 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3541 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     3601 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     3661 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
     3721 tcactata
//
