LOCUS       sequence-8531-ex        5168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5168)
  AUTHORS   Thorpe CJ, Weidinger G, Moon RT
  TITLE     Wnt/beta-catenin regulation of the Sp1-related transcription factor 
            sp5l promotes tail development in zebrafish.
  JOURNAL   Development. 2005 Apr. 132(8):1763-72.
  PUBMED    15772132
REFERENCE   2  (bases 1 to 5168)
  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: 
            "Development. 2005 Apr. 132(8):1763-72."
COMMENT     Sequence Label: ZE38 pCS2P+ sp5l ORF
FEATURES             Location/Qualifiers
     source          1..5168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             411..440
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             414..443
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1193..1212)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1194..1212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1217..1351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1241..1260)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1295..1314
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1464..1484)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1464..1482)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1503..1519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1503..1519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1516..1538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1527..1543
                     /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(1551..1557,1558..1575,1576..1581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1596..1617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1734..1751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1905..2493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1985..2004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2664..3455,3456..3524))
                     /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     3287..3306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3525..3629)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3655..4110)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3792..3813)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4004..4023
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4184..5168
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcc caccatggct gcattaacct
      121 tatccagagc ggacaatttc ctccacaatt tcttacagga ccgcactccc agctcatctc
      181 cagaaagtgg tccaaacacc ctctcctttc tggccaccac atgcagtcag gcttggcagg
      241 aaggctcgca gctcccttac gagggtccgg tgggctccgc atccagcatg ttccagctct
      301 ggagcaacga cgtggccccg aactccagct taagcgctca ccagatgacc ttcaccgtgc
      361 ccaaaatgca gtttcccagc cacatgcaac ccactctggg ctcgcactcc catcaccacc
      421 atcaccacca tcatcaccac cacgagcttc ccctcactcc tccagcggag cctccgtctg
      481 catattcatt cgagctttcc ccggttaaaa tgcaagggaa tgcgccatac tacacgcagc
      541 acaacgccgt cagtcaaaac tttccaagct tcctccaaaa cccctcagga agagctcacc
      601 tgcctggagg tcacgttgag gatggccagc agtggtggag tctcccccag ggtaacagcg
      661 ctccctccgg gcaccctttc tccttgggac ggcagctggt tttgggccat cagcctcaaa
      721 ttgcagcact tctgcaggga acctccaaag gtctgttgag ctctactcgt cgctgtcgcc
      781 gatgcaaatg cccaaactgc cagtccacag gaaacggagg tgctgcattg gagtttggaa
      841 agaaaagact acacatttgt cacattccag attgtgggaa ggtttataaa aaaacctctc
      901 acctcaaagc gcacctgcgc tggcatgctg gtgaacggcc cttcatttgc aactggctct
      961 tctgcgggaa gagcttcacg cgttcagatg agctgcagag gcatctccgt acacacaccg
     1021 gggagaagcg tttcggctgt cagcagtgtg gaaaacggtt tatgcggagc gatcatctct
     1081 ccaagcatgt gaagacccat cagagccgga agagccggtc cagtcagcct cctcacagcg
     1141 ggactgatgc tctgctgagc aacatcaaga gagagtaatc tagattctgc agccctatag
     1201 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     1261 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1321 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1381 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg
     1441 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     1501 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2341 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2941 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3541 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3601 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     3661 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3721 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3781 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3841 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3901 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3961 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4021 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4081 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     4141 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccata gccaattcaa
     4201 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa
     4261 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa
     4321 tatggcggac ttggcaccat gccaattcaa tatggcggac ttggcactgt gccaactggg
     4381 gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc
     4441 gccatattga attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca
     4501 atatataggc aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac
     4561 tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata
     4621 gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc
     4681 cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg
     4741 cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc
     4801 ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct
     4861 cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc
     4921 acttggcagt acatcaatat ctattaatag taacttggca agtacattac tattggaagg
     4981 acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg
     5041 gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc
     5101 cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt
     5161 tagggaac
//
