LOCUS       sequence-80478-e        3734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  actinodin 4 and4 antisense in situ probe linearize w/ AatII SP6 pol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3734)
  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 3734)
  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-and4 in situ probe
FEATURES             Location/Qualifiers
     source          1..3734
                     /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(859..877)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(860..877)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(895..911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(895..911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(908..930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    919..935
                     /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(943..949,950..967,968..973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    988..1009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1126..1143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1297..1885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1377..1396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2056..2847,2848..2916))
                     /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     2679..2698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2917..3021)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3099..3554)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3236..3257)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3448..3467
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3680..3702
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3694..3711
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3695..3711
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3718..3734,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3718..3734,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tcgcacagaa ctcttacctg tccattcatc accgacagat catcctcaga agcagagcgc
      121 gcttaatgac gagtgtgtgt gtgtgtgagg gtcagaggtc atgatgtctg cggtgtgtat
      181 gctgtccgtc ctgctgctga tctgcggtca gctggaggcg aaatccctga tggagatcct
      241 cagcccggat ggagcgctca gtctggactc cagcaaggct catgagtttc tgtccagttc
      301 tcgacccaag cggagcctgg acccacgctg gcaccggcag agcccagact tccaggccta
      361 ctacaaatac tacagcagca tcgggcacac agagggcctg tatgaggtgg accggatccg
      421 catgcagtac cagcagatgc ggcacctgga gcagctccac ggccccgatg caccctactt
      481 ccagaaccgg ctgggccgac cggcgcttcc tgcgcttccc aaatgtgatc ctgcaacaga
      541 caagggctgc agagtgtccc cgcccccagc catagcccca ccccccaccg cagtgaaggg
      601 ccctgtggct ccgccccctg ctgccgtgaa gggccccgtg gctccgcccc tcagtcaggc
      661 agatgtggtg tatctgtgca atgctaaaga cccgctctgc aggccgcaca tcgtctacat
      721 gcccgcgggg gccgtcccgg tgctgtgcga cccacgctac catcccaact gcaagctgaa
      781 tcactagtga attcgcggcc gcctgcaggt cgaccatatg ggagagctcc caacgcgttg
      841 gatgcatagc ttgagtattc tatagtgtca cctaaatagc ttggcgtaat catggtcata
      901 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      961 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1021 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1081 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1921 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1981 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2041 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2101 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2161 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2221 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2281 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2341 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2401 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2461 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2521 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2581 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2641 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2701 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2761 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2821 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2881 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2941 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3001 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgatg cggtgtgaaa
     3061 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     3121 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3181 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3241 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3301 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3361 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3421 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3481 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3541 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     3601 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     3661 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
     3721 tacgactcac tata
//
