LOCUS       62056_21230        5548 bp DNA     circular SYN 26-APR-2021
DEFINITION  Cloning vector pTol2-HS-GT2MP-EGFP, complete sequence.
ACCESSION   MW698954
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5548)
  AUTHORS   Lu C, Fan C, Wang J, Zhao Y.
  TITLE     Identification of muscle highly expression proximal cis-regulatory 
            element in zebrafish
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5548)
  AUTHORS   Lu C, Fan C, Wang J, Zhao Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAR-2021) College of Fisheries and Life Science, 
            ShangHai Ocean University, No.999, Huchenghuan Rd, Nanhui, Shanghai 
            201306, China
REFERENCE   3  (bases 1 to 5548)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-MAR-2021) College of Fisheries and Life Science, ShangHai Ocean 
            University, No.999, Huchenghuan Rd, Nanhui, Shanghai 201306, China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        411..515
                     /label=AmpR promoter
     misc_feature    700..1077
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene;
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     rep_origin      1546..2134
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2422..2443
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2458..2488
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2496..2512
                     /label=lac operator
                     /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)."
     primer_bind     2520..2536
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2557..2575
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2663..2868
                     /label=tol2-L
                     /note="tol2-L"
     misc_feature    2875..2946
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus
                     (Farrell et al., 2002)"
     CDS             3814..4527
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    4575..4799
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4813..4904
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     misc_feature    4919..4990
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus
                     (Farrell et al., 2002)"
     repeat_region   5112..5261
                     /label=tol2-R
                     /note="tol2-R"
     promoter        complement(5292..5310)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5320..5336)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5478..5548
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
       61 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
      121 cacctcgacc gcaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
      181 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
      241 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
      301 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
      361 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
      421 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      481 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      541 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
      601 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
      661 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
      721 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
      781 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
      841 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
      901 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
      961 ttttgcacaa catgggggat catgtaactc gccttgatcg ttggaaccgg agctgaatga
     1021 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1081 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1141 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1201 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1261 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1321 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1381 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1441 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1681 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2221 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2281 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2341 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2401 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     2461 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     2521 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     2581 aaagctggag ctccaccgcg gtggcggccg ctctagaaac tagtggatct gctgggcttg
     2641 ctgaaggtag ggggtcaaga accagaggtg taaagtactt gagtaatttt acttgattac
     2701 tgtacttaag tattattttt ggggattttt actttacttg agtacaatta aaaatcaata
     2761 cttttacttt tacttaatta cattttctta gaaaaaaaag tactttttac tccttacaat
     2821 tttatttaca gtcaaaaagt acttattttt tggagatcac ttgggccctg accccatctt
     2881 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     2941 cctctctccc ttgatgattt atatataatt tcttgtacaa tatattgtct caaactgcta
     3001 cagctgcgct gaatgatgag tctctcagct ctctggatgt ggagcagtag gccgacagtc
     3061 gccgtggcat ttcggaaagc atgctgtccg agccaatggc agtcagcgcg ctctgctatt
     3121 ggttcccggg gcgctcactg ccagctcgtg tccccgccca tgttcgtaag atatggaatc
     3181 tactggcgcc agttccgaca gtacacaggc acaattcatt aatgagactt ctctccgctt
     3241 cagacagacg cagagtttta gggagacttt aacaatcggg ctgtggacaa tttaaaccag
     3301 tggcgaatta cgaacgtcaa caggcatctt gaggattaac attctttgcg caggactaac
     3361 acgggaaaaa taaacgcagg attggagtgc tgaaatgcaa ctttgcgccg tgagtacttc
     3421 ccggtagtta tttgaaattg cgagcaattt aattgagcga tttaattgat tgactacaaa
     3481 agttagccta cttatattaa ctgaggcgtc gtcgtgtgaa ttaagatctg tcttgcactg
     3541 tgtttaacgt caacactgag atgcttctat ccgttattct cttacaggtg tccctggcca
     3601 cccttgaatg caaagaagca ggacctctac actccttcaa aaataaaagc atgctcagaa
     3661 agtaaacaga gcatcgccac ctgaagcatt aagctaacga cagatatttt aataatctaa
     3721 cggactatag tggtgctttc gggtctgtag tgtcaagtaa acttttccaa gcattttcta
     3781 agcgcggaca cttgagccgc caccatgtct agagtgagca agggcgagga gctgttcacc
     3841 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3901 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3961 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     4021 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4081 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4141 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4201 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4261 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     4321 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4381 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     4441 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4501 actctcggca tggacgagct gtacaagtaa agcggccgcc accgcggtgg agctcgaatt
     4561 aattcatcga tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4621 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4681 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4741 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4801 ttctgagaat tcaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
     4861 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat gatgatccca
     4921 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
     4981 tccctcccta aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     5041 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct
     5101 agatcagatc taatactcaa gtacaatttt aatggagtac ttttttactt ttactcaagt
     5161 aagattctag ccagatactt ttacttttaa ttgagtaaaa ttttccctaa gtacttgtac
     5221 tttcacttga gtaaaatttt tgagtacttt ttacacctct gtcaagaacc atatgccggt
     5281 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     5341 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5401 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5461 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5521 tacgcgcagc gtgaccgcta cacttgcc
//
