LOCUS       sequence-101757-        6729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis of tol1 transposase codon modified for zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6729)
  AUTHORS   Horstick EJ, Jordan DC, Bergeron SA, Tabor KM, Serpe M, Feldman B, 
            Burgess HA
  TITLE     Increased functional protein expression using nucleotide sequence 
            features enriched in highly expressed genes in zebrafish.
  JOURNAL   Nucleic Acids Res. 2015 Jan 27. pii: gkv035.
  PUBMED    25628360
REFERENCE   2  (bases 1 to 6729)
  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: "Nucleic 
            Acids Res. 2015 Jan 27. pii: gkv035."
COMMENT     Sequence Label: pCS2-Tol1.zf1
FEATURES             Location/Qualifiers
     source          1..6729
                     /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"
     polyA_signal    2778..2912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2802..2821)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2856..2875
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3025..3045)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3025..3043)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3064..3080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3064..3080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3077..3099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3088..3104
                     /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(3112..3118,3119..3136,3137..3142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3157..3178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3295..3312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3466..4054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3546..3565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4225..5016,5017..5085))
                     /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     4848..4867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5086..5190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5216..5671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5353..5374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5565..5584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5745..6729
                     /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 ttcgaattcg agctcggtac ctcgcgaatg
      121 catctagata cccaatgcca tcatggaaaa aaaacgatca aaaccctcag gcgcgcaatt
      181 cagaaagaag agaaaggagg aggaggagaa gagagacaag gagaagggcg ctctgctgag
      241 atactttgga tctagcacca ctgctcagga tgagaccagc acctctctgc ctgccatctc
      301 ttctgccact gtgactgtgt ctcctcctca ggatgagctg cctagcacct cttctgccac
      361 ccatgtggtg cctcagctgc tgcctgagca gtcttttgac tctgaggctg aggatgtggt
      421 gcctagcacc agcacccagc tggagacctc tgagatgcct ggagatgaga cccctctgac
      481 cccaactgct gaggaccagc ctctgccaac tgaccctgcc aagtggcctt ctcctctgac
      541 tgacagaatc agaatggagc tggtgagaag aggaccttct agcatccctc ctgactttgt
      601 gttcccaaga aatgactctg atggacgctc ttgccaccac cactacttca gaaagaccct
      661 ggtgtctgga gagaagattg ccagaacctg gctgatgtac agcaaggtga agaactctct
      721 gttctgcttc tgctgcaagc tgttcagcaa caagaacatc aacctgacca cctctggaac
      781 tgccaactgg aagcatgcta gcacctacct gactgctcat gagaagtctc ctgagcacct
      841 gaactgcatg aaggcttgga aggagctgtc tggaagaatc cgctctggaa agaccattga
      901 caagcaggag atggctctgc tggaggagga gagagtgaga tggagagctg tgctgaccag
      961 actgattgcc attgtgcagt ctctggctgt gagaaacctg gctctgagag gacacactga
     1021 gaccctgttc acctctagca atggaaactt cctgaaggag gtggagctga tggccagatt
     1081 tgacccaatc atgaaggacc acctgaacag agtgctgaga ggaactgctt ctcacaactc
     1141 ttacattgga caccatgtgc agaatgagct gattgacctg ctgtctagca agatcctgtc
     1201 tgccattgtg gatgacatca agaaggccaa gtacttcagc atcatcctgg actgcaccct
     1261 ggacatctct cacactgagc agctgtctgt gatcatcaga gtggtgtctc tgatggagaa
     1321 gcctcagatc agagagcact tcatgggatt cctggaggct gaggagagca ctggacagca
     1381 cctggctagc atgatcctga acagactgga ggagctggga atctcttttg aggattgcag
     1441 aggacagtct tatgacaatg gagccaacat gaagggaaag aacaagggag tgcaggccag
     1501 actgctggag aagaacccac gcgctctgtt cctgccttgt ggcgctcaca ccctgaacct
     1561 ggtggtgtgt gatgctgcca agcgctctgt ggatgccatg tcttactttg gagtgctcca
     1621 gaagctgtac accctgttct ctgcttctgc tcagagatgg gccatcctga agtctcaggt
     1681 gagcatcacc ctgaagtctt ggactgagac cagatgggag agcaagatca agagcattga
     1741 gccaatgaga taccagggag ctgctgtgag agaggctctg attgaggtga gagacaagac
     1801 caaggaccct gtgatcaagg ctgaggctca gtctctgtct gaggaggtgg gatcttacag
     1861 attcaacatc tgcactgtgg tgtggcatga catcctgagc accatcaagc atgtgagcaa
     1921 gctgatgcag tctccaaaca tgcatgtgga cctggctgtg tctctgctga agaagactga
     1981 gcagtctctc cagtcttaca gagccaatgg atttgtgaat gctcagatgg ctgccaagga
     2041 gatgtgcaag gagatgaatg tggaggccat cctgaagcag aagagaatca gaagcaccaa
     2101 gtgccagttc tcttatgagt ctcatgatga gcctttctct gatgctctga agaagctgga
     2161 ggtggagttc ttcaatgtgg tggtggatga ggctctgtct gccattgctg agagattcag
     2221 caccctggag gtggtgcaga acagatttgg agtgctgacc aacttccctt ctctgggaga
     2281 tgaggagctg actgagcagt gtgaggctct gggaaacatc ctgcactttg agaagaactg
     2341 ggacctggac agcagagagc tggtgcagga gatcaagaac ctgccaaacc tgcctagcac
     2401 caccccttct ctgctggagc tgatctcttt catgtctgac aaggacctgt ctgaaatcta
     2461 cccaaacttc tggactgctc tgagaattgc tctgaccctg cctgtgactg tggctcaggc
     2521 tgagcgctct ttcagcaagc tgaagctgat caagtcttac ctgagaagca ccatgtctca
     2581 ggagagactg accaacctgg ctgtggtgag catcaaccac tctgtgggag agcaaatctc
     2641 ttatgatgat gtgattgatg agtttgctag cagaaaggcc agaaaggtga gattctaaaa
     2701 ttgggtcatc ggatcccggg cccgtcgact gcagaggcct ctcgagcctc tagaactata
     2761 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     2821 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2881 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2941 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt
     3001 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3061 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3121 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3901 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5221 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5281 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5341 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5401 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5461 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5521 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5581 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5641 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     5701 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca
     5761 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     5821 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
     5881 atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg
     5941 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
     6001 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
     6061 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
     6121 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
     6181 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
     6241 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
     6301 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
     6361 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
     6421 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
     6481 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
     6541 tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
     6601 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
     6661 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
     6721 ttagggaac
//
