LOCUS       sequence-212865-        4543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains zebrafish sod1 gene in the pCS2+ expression vector. It has 
            the SP6 RNA polymerase site for in vitro transcription..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4543)
  AUTHORS   Zhou W, Cao L, Jeffries J, Zhu X, Staiger CJ, Deng Q
  TITLE     Neutrophil-specific knockout demonstrates a role for mitochondria in
            regulating neutrophil motility in zebrafish.
  JOURNAL   Dis Model Mech. 2018 Mar 28;11(3). pii: dmm.033027. doi: 
            10.1242/dmm.033027.
  PUBMED    29590639
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"; journalName: "Dis Model 
            Mech."; date: "2018-03-28"; volume: "11(3). pii: dmm.033027. doi"; 
            pages: " 10.1242/dmm.033027"
COMMENT     Sequence Label: pCS2-sod1
FEATURES             Location/Qualifiers
     source          1..4543
                     /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"
     regulatory      87..96
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    595..729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(619..638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     673..692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(842..862)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(842..860)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(881..897)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(881..897)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(894..916)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    905..921
                     /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(929..935,936..953,954..959))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    974..995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1112..1129)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1283..1871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1363..1382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2042..2833,2834..2902))
                     /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     2665..2684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2903..3007)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3033..3488)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3170..3191)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3382..3401
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3559..4543
                     /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 atcctcgcca ccatggtgaa caaggccgtt tgtgtgctta
      121 aaggcaccgg tgaagtgacc ggcaccgtct atttcaatca agagggtgaa aagaagccag
      181 tgaaggtgac tggtgaaatt actggcctta ctccaggaaa acatggtttc cacgtccatg
      241 cttttggtga caacacaaac ggctgcatca gtgcaggtcc gcacttcaac cctcatgaca
      301 aaactcatgg tgggccaacc gatagtgtga gacacgtcgg agacctgggt aatgtgaccg
      361 ctgatgccag tggtgttgca aaaattgaaa tcgaggatgc aatgctaact ttgtcaggcc
      421 aacattctat tattgggagg accatggtga ttcatgagaa ggaggatgac ttggggaagg
      481 gtggcaatga ggaaagtctt aaaactggca acgctggtgg tcgtctggcc tgtggagtga
      541 tcggcatcac tcagtgactc gagcctctag aactatagtg agtcgtatta cgtagatcca
      601 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
      661 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
      721 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
      781 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
      841 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
      901 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
      961 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1021 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1081 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1141 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1201 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1261 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1321 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1381 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1441 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1501 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1561 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1621 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1681 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1741 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1801 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1861 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1921 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1981 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2041 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2101 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2161 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2221 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2281 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2341 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2401 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2461 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2521 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2581 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2641 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2701 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2761 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2821 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2881 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     2941 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3001 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     3061 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3121 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3181 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3241 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3301 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3361 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3421 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3481 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     3541 ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa
     3601 ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa
     3661 ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa
     3721 ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca
     3781 agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca
     3841 ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc
     3901 cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     3961 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     4021 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     4081 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     4141 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     4201 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccgttctca
     4261 cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt
     4321 aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc
     4381 ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg
     4441 tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg
     4501 tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
