LOCUS       sequence-174299-        4114 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  To make in situ hybridization probe for Xenopus amylase. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4114)
  TITLE     Spagnoli lab plasmids
REFERENCE   2  (bases 1 to 4114)
  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"
COMMENT     Sequence Label: x.Amylase
FEATURES             Location/Qualifiers
     source          1..4114
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             227..1021
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             227..1021
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(281..300)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     891..910
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(1271..2062,2063..2131))
                     /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     1894..1913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2255..2843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2744..2763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2997..3014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3131..3152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3167..3172,3173..3190,3191..3197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3205..3221
                     /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)."
     primer_bind     3210..3232
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3229..3245
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3229..3245
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3266..3284
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4003..4022)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4004..4022)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4029..4046)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4029..4045)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4038..4060)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
       61 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
      121 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
      181 caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
      241 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
      301 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
      361 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
      421 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
      481 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
      541 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
      601 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
      661 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
      721 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
      781 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
      841 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
      901 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
      961 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     1021 aattattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1081 ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1141 ttttctaaat acattcaaat atgtatccgc tcatgagatt atcaaaaagg atcttcacct
     1201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1261 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1321 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1381 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     1441 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1501 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1561 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1621 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1681 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1741 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1801 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     1861 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     1921 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1981 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2041 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2101 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2161 tttatcaggg ttattgtctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2221 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2281 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2341 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2401 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2461 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2521 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2581 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2641 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2701 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2761 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2821 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2881 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2941 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3001 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     3061 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     3121 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     3181 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     3241 atgaccatga ttacgccaag ctcagaatta accctcacta aagggactag tcctgcaggt
     3301 ttaaacgaat tcgcccttgc catcttttct ccattccatt tgcggatgac tcccccaagc
     3361 ttggcaccat atttgaattc agtaactcgt ccaactccaa agtattcgtt gacagaaatt
     3421 gcttcacccc caagatcaat tacctcttgg aagatgaaag gtcgagctcc tgcagggaac
     3481 cattttgtgt tcagattatt gagtctgtcg gaaatagcct ttaggtctcc aggccacata
     3541 tgtttagcag catccagtct gaatccagca accccaatgt taattagatt gttcatgtat
     3601 tcggcaatct tcccacgaac atagtctttc tccattgcca aatccaagag accaacaaga
     3661 cgacagtttc ggacctggtt agcatcccca taattttcta tttcaccact gccagtcctg
     3721 cactttccat cattaaaatc caagccggaa taggggactg ggaaatccct ggatcctgca
     3781 ttgaagtagc ttccacaagt tgaatgagtg ccagcacctc cacctgagcc acacatatgg
     3841 ttgattatgg catccacata aatatatacc ccaacattgt tacatcttgt caccatgtct
     3901 ctaaactgtt gttcattccc agaacgagtg cagagcttat aacttatagg ttggtaactc
     3961 tcccaccaaa agggcgaatt cgcggccgct aaattcaatt cgccctatag tgagtcgtat
     4021 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4081 cttaatcgcc ttgcagcaca tccccctttc gcca
//
