LOCUS       sequence-12556-e        4839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T/A Cloning Vector for dsRNA generation and the generation of sense 
            and anti-sense probes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4839)
  AUTHORS   Collins JJ, Hou X, Romanova EV, Lambrus BG, Miller CM, Saberi A, 
            Sweedler JV, Newmark PA
  TITLE     Genome-wide analyses reveal a role for peptide hormones in planarian
            germline development.
  JOURNAL   PLoS Biol. 2010  . 8(10):e1000509.
  PUBMED    20967238
REFERENCE   2  (bases 1 to 4839)
  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: "PLoS Biol. 
            2010  . 8(10):e1000509."
COMMENT     Sequence Label: pJC53.2
FEATURES             Location/Qualifiers
     source          1..4839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..49)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        63..81
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        85..103
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        237..339
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             340..1023
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(406..425)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1343..1648
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1527..1546
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        complement(1719..1737)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1720..1737)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1741..1759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1773..1820
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1832..1849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2003..2591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2083..2102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2762..3553,3554..3622))
                     /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     3385..3404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(3640..4434)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3751..3770)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4361..4380
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagg cgcgccggat
       61 cctaatacga ctcactatag ggagaattaa ccctcactaa agggagacct taggtcgcgg
      121 ccgctttgcg ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct
      181 ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat
      241 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
      301 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
      361 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
      421 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
      481 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
      541 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
      601 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
      661 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
      721 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
      781 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
      841 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
      901 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
      961 ttacaacagt actgcgatga gtggcagggg gggcgtaaac gccgcgtgga tccggcttac
     1021 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata
     1081 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt
     1141 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt
     1201 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg
     1261 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct
     1321 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg
     1381 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
     1441 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     1501 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     1561 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     1621 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc
     1681 aggtcgatac agtagaaact cgaggaccta aggtcacttc tatagtgtca cctaaatctc
     1741 cctatagtga gtcgtattag gatccggcgc gcctagcata accccttggg gcctctaaac
     1801 gggtcttgag gggttttttg acgcgtgctt cctcgctcac tgactcgctg cgctcggtcg
     1861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2401 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2821 tagttgcctg gctccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3601 cacggaaatg ttgaatactc atactcttcc tttttcaatt cagaagaact cgtcaagaag
     3661 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     3721 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     3781 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     3841 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     3901 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     3961 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     4021 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     4081 atcagccatg atggatactt tctcggcagg agcaaggtgg gatgacagga gatcctgccc
     4141 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     4201 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     4261 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     4321 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     4381 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     4441 cgatcctcat cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg
     4501 caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc
     4561 tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag
     4621 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca
     4681 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg
     4741 cttcctttag cagcccttgc gccctgaatt ttgttaaaat tcgcgttaaa tttttgttaa
     4801 atcagctcat tttttaacca ataggccgaa atcgacgcg
//
