LOCUS       sequence-177131-        6904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in situ hybridization, linearize with Sac I use T7 polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6904)
  AUTHORS   Rubinstein AL, Lee D, Luo R, Henion PD, Halpern ME
  TITLE     Genes dependent on zebrafish cyclops function identified by AFLP 
            differential gene expression screen.
  JOURNAL   Genesis. 2000 Jan;26(1):86-97.
  PUBMED    10660676
REFERENCE   2  (bases 1 to 6904)
  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: "Genesis."; 
            date: "2000-01"; volume: "26(1)"; pages: "86-97"
COMMENT     Sequence Label: crestin
FEATURES             Location/Qualifiers
     source          1..6904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3953..3969)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3953..3969)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4006..4024)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4045..4061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4045..4061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4058..4080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4069..4085
                     /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(4093..4099,4100..4117,4118..4123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4138..4159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4276..4293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4447..5035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4527..4546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5206..5997,5998..6066))
                     /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     5829..5848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6067..6171)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6198..6653
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6285..6304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6495..6516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6780..6802
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6794..6811
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6795..6811
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6818..6837
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6818..6836
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gatttattgg gctacactac gataaatgat actaactacg taataagcaa acaacaaaca
       61 aacacacaca ttcacacata cacacaaagg cagttcagag gatgcaaaat gagttgacaa
      121 cgtcccaaat taattctaat acttcttgca agatcttaga ataacacctg agaaaccaca
      181 aaagcagaga tatggcttat ctttaactgc ttaagatcaa tctgcacaag atcagcaaga
      241 gacaacatta gtttgtgtta cttgcgccct ttttgctgaa gtcgaatctc cctcgccgac
      301 tggccttggg gaaacacggt gctcctgatt gggtggtttc tcgcttcaat gacgtctttg
      361 ggattcacgc gctcgtgaat ggctgttgtc ctgggttacc gaggagacgg acagctgaga
      421 gtcggtttgc ggaagttcat gcagcttcgg gagtcccgaa acttaagagg tttgcgtgga
      481 agttcgtagt gacccaatcg cgtggcggcg cggtgtccgg gcaggcgaaa gggggcagag
      541 agcaaaagca tgcgcaagct agctgatgca aaaagcaaag tttctttgtc gcagggtgtt
      601 tttaagtgac ctggtctggt tcgtccccat ggcactgtct gccaatgagc agttgccatg
      661 aagggcgggc cacaccccgt actgacgtag ggaaggtcaa taattaacag actgcatgta
      721 gatttcaagt ggtaaaactc ttgcaagatt gaaactatct tacattaaac gttataagat
      781 taatactgtt tgtactacta gtttttcatt tacaccaacg tattgcaaat gtcactagct
      841 ttcgttcaat accagacatc atacatttca gacacataca gaacatttta ctctgttata
      901 agggttaaaa caacacattg attaacctgg ttgatttgca ataaacagca tatgaggaac
      961 attatggcat attattcaaa gatacatctg ccttagttct aacattttaa gagattgtct
     1021 ttcattcata tgcattaatg tttctagaga tctctttgtc tctacaaaat tgtgcttcta
     1081 gcaggatgtg cccaaggcga gcacatgacc aggagtcaat caaaagtctc gtgaaatggt
     1141 ttccgctgaa gatcctgtaa gggccattgt gttttaattt aactgattta attaatcatt
     1201 ggctagttag ccgaagggta ccagaacatc tgcggagaag tactggtcag aggagcccaa
     1261 aatcaattgg cgctctgtta tcctgatggt ataagccctc aagctagttc aatgaatgag
     1321 accattggag atgcccaaga tttcagattc aacttgtgtt gaccactgaa aaaggccgat
     1381 gagttcctac agtccccctt tgactggtga tgttcccggt gtgaacattg gcaggcactg
     1441 gaacaaacag gcagaagcag agagaacagg caaacacaag acacaaacaa catcatcact
     1501 aactgaatct tggtgcaggg tttaggaact ttggagtgct cgggttagca atgctcaagg
     1561 tgtagaagag catagtttag tatgatcaga atttaggctt ttaagactag ccgaggtatt
     1621 gaaggttcaa cgaagttgta taggccttat ctcaatcagt aaagaattta atttttggtc
     1681 agcgttatcg catctaagca tgcgcgagat gtggggcgtg cacatttgaa caacttttgg
     1741 atggccttta ctcgctcgtg gaggaagagg gcaagtgtca aagtcaacgt ataccctaca
     1801 atgctatttt gacgagagtc aaaccaataa atttcattct ttaccctcaa tccctttttc
     1861 gttctggatt gaagatgaag aaatgtggta aacaaatctt ttgagttttc gcctaaaacg
     1921 tgtcgtcaga caggtgccac aacgttaaat taccaataca gcgatcacac ctttaaggca
     1981 ttgatccgtg aggcttggtg aaaggagtca atcctaattc tgagctctgg acttaaacat
     2041 catctgtggg ggttccctga tgaggcaaga gaaattaaaa gacagagaga aagagaaaga
     2101 aaagaacact gagacataag gacattgagt gaggttaaga caaacaaatg tcacatgtaa
     2161 tagctcctct ctactagaga ttggtcctag tagtgcagtg gaggggagag tgcatgtatt
     2221 gggtttagga aacatagcta gggctaggtg tcttcctaaa atggcacaaa tgacaaaaac
     2281 aggttagcgt taaaataaga gtttaggata gtgggtctgt gtcagcaggt tggtccctcc
     2341 ccctttctgg tcacttggag ttgagggact ggagttggca tggcacaggt aggatttgga
     2401 ttgattctga gttcagctgc gtggcaaacg atcaagacag cactcttagg aattggaagg
     2461 gtttcatcca caagtgtttg tgtgatgcag tggtgacctg aggcatcagg tgttgagtta
     2521 gcgtctaact ctggtgttca ggtacagtct attgagtctt tgaagttatg gtgcttacag
     2581 ttctctgtaa gtgaggtgaa gaggtgctgt ttagtgaggt cgttgcttgt caggcagaat
     2641 caggttttcc tttcctttcc cacttccgat ccgcttccct tcgttggaag aactctttca
     2701 ggatcatttt catcagttct tgagggtcaa aacatggtga tgttgtccct tgtggggatt
     2761 cagttggagc tcgatcagcg tggaatcttt gtgagttctt tcgttgacct cttgggctgg
     2821 ttgtatcaag gtgatgttgc tttaccttgg aagtaccact tgattcccac gagcttgtcc
     2881 ccctttggtt tacaaatgag gtagacctgt tccacggtct ctcccggtga cgttcaggtg
     2941 agtgttgtcg tccagacagt cctttccagc gacggttcca ctgtttgggt ctggtatcag
     3001 catgacagtc tcgttgtctg ttgtaagacg acacattcca ctcactgtga ggtagtctag
     3061 cattgtctgg gccttgggca ccctcaaggg ccagatcttg actttgagtg ttgaagtcat
     3121 aaacagcagg gattttggca ctcttctctg aagccgtctt ttgtttgacg tatgctttat
     3181 gtgccaaatc tcgtatttgc tgagtgttca tggtgtgggg gcatgcaaga accccaagat
     3241 ggtggcttac cccaggatgg aggtttctca ggaagaggat tttgaagttc aggtcctctt
     3301 ccatctcagg ttcattgcgg gtgccgaagt aggctcgtct gagtctgctg tagtaggctt
     3361 gagaagactc atggcgactt tgtttcgtct ccagggcagc tacaagtcct tgttcagact
     3421 cgatgtcatc aaactctctg atgagggctt ctcggagcag gaggtagtca gtctttgtgt
     3481 gagccggctg acggtccagg aagctgcgca cttcagggct ggaggttgct cggagcaaat
     3541 acagtctatc tttgtcagtg acattgggtc gcatttccag gtgaaagtca atgtcctgca
     3601 gataggcgtg aacatcttgg ctacctggca aattcggcgt gaacttgccc aggttcttga
     3661 ctagcttgtc aaggtctttg aggtccaggc cacagggtga tacgtggctg tctggaacag
     3721 aaggagatgg tgagggtttt ggcagtgacc cctcagcttt ctcattgtgt gtcagttcat
     3781 atgcatgctt gagttcatct ttaatgctgt tggactcttc tttgatgtag tccagctgct
     3841 gcgttagtcg tctgacctca tgtctagctt cactcaggtg agtttcgagg gctttgattc
     3901 tgctgtcctt gtctttagag tcagcctcgt gccgaattcc tgcagcccgg gggatccact
     3961 agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     4021 aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4081 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4141 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4201 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4261 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4321 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4381 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4441 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4501 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4561 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4621 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4681 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4741 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4801 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4861 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4921 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4981 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5041 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5101 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5161 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5221 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5281 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5341 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5401 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5461 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5521 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5581 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5641 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5701 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5761 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5821 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5881 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5941 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6001 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6061 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6121 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6181 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6241 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6301 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6361 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6421 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6481 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6541 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     6601 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttgccattc
     6661 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6721 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6781 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga
     6841 attgggtacc gggccccccc tcgagttttt tttttttttt tttttttttt tttttgagta
     6901 caaa
//
