LOCUS       sequence-80476-e        4543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  actinodin 2 and2 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 4543)
  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 4543)
  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-and2 in situ probe
FEATURES             Location/Qualifiers
     source          1..4543
                     /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(1668..1686)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1669..1686)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1704..1720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1704..1720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1717..1739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1728..1744
                     /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(1752..1758,1759..1776,1777..1782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1797..1818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1935..1952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2106..2694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2186..2205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2865..3656,3657..3725))
                     /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     3488..3507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3726..3830)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3908..4363)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4045..4066)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4257..4276
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4489..4511
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4503..4520
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4504..4520
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4527..4543,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4527..4543,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 tatggccaga ctcattaaga tctttgcaac agctcttgtg atcgtcttca tgtcagactt
      121 cttgagtgct ggtcctgtaa agcgtaatga agaagtggat ggtgcatctg aggtggaaac
      181 agacagcaag tcacatatcc gcaaaaggag gaatattgcc ttctacagaa gtcaacctga
      241 tttctggggc tggtataaat tcttcatgga gaccaacaac caggagggaa ttgaggactt
      301 ggatcgaatg taccgtgttt acctgcagaa caagcacaga gtagaagagg gtgccaattt
      361 caaccattac ctcactcact tgagtcaaat ttacaaaacc tgtgccaact ctgatgaccc
      421 tgattgcatt gcagagtcca ccagcaagcc caaagctaat attgtcatgc ctttacctgt
      481 gaggcaagct actgtggcag tgtgcaaccc ttaccttgac ccttactgcc tctacgccat
      541 cagaccaaaa gctgcagaag agccttcacc tgccccagca aaggtccctg ccccaattct
      601 gtcacctctg ctcccgctgc ccctgaaagc acctctagcc cattattact atgctccagt
      661 catggagcca tttttgtcag ctgagcagag agcagagctg ttgcgcatct gcaacccttc
      721 agacgcagag tgtctgcaat acacttgcga gctgcttacg gctacagacc cgcactcggc
      781 cctctgcctt cttactctca ccttggatgt gatcctacca aagatcctta ctgtcagccc
      841 aagcttgtgg caagatctcc ctcaggtttg taccacctgt acccttcctg taatcccgcc
      901 actgaccctc tttgtgttgc caatgttgtt gcacctgcag ctcaaaccgc tgagagtgca
      961 gaagccccaa aagacaagct ctgcaaccct ttgtttgacg aaggctgtaa cccccttaca
     1021 gctgccaagc tagcggctct caataaacct gtcctggaat atgctccaag agatgagcct
     1081 gcacctctaa acctggcctg tgacccgcgt tacgacccct attgtctgat aggtggatcc
     1141 gctgctctca ggaagccccc tccagttctc ccagagttcc agacccgtca caatctgggt
     1201 gtccgtggaa agaccaagga aggttatgac tgctatatgt tttacgataa ggactgcacc
     1261 ccagtggaga atcaggatct gaggtccagt gctgccagtt ccaagccaga ctgccaccca
     1321 tttgacccca actgtgggag gtttgccgct cagccatcca ctggagcaga gccagctaaa
     1381 acagtcaaga atggcatcat tgagccccat cccgactgtg acccagaaat tgattacaac
     1441 tgtcgcctgc gccggtcaga atctgcaggt gatgaaccag cccagccaga acaaggtcat
     1501 ggcaaacctg aacacgtggc tcccgagtat cctgttccaa gatttgagga cttcctcaga
     1561 gcatacgtgg gtggctacaa gaaatgaaat cactagtgaa ttcgcggccg cctgcaggtc
     1621 gaccatatgg gagagctccc aacgcgttgg atgcatagct tgagtattct atagtgtcac
     1681 ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1741 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1801 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1861 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2521 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2761 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2821 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2881 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2941 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3001 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3061 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3121 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3181 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3241 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3301 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3361 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3421 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3481 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3541 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3601 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3661 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3721 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3781 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3841 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     3901 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3961 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4021 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4081 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4141 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4201 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4261 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4321 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     4381 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     4441 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4501 gttgtaaaac gacggccagt gaattgtaat acgactcact ata
//
