LOCUS       sequence-148104-        6925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector bearing an NheI restriction site for the two-step cloning 
            method in Wang and Szaro, Dev Biol, 2015.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6925)
  AUTHORS   Wang C, Szaro BG
  TITLE     A method for using direct injection of plasmid DNA to study 
            cis-regulatory element activity in F0 Xenopus embryos and tadpoles.
  JOURNAL   Dev Biol. 2015 Feb 1;398(1):11-23. doi: 10.1016/j.ydbio.2014.11.010.
            Epub 2014 Nov 20.
  PUBMED    25448690
REFERENCE   2  (bases 1 to 6925)
  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"; doi: 
            "10.1016/j.ydbio.2014.11.010"; journalName: "Dev Biol"; date: 
            "2015-02-1- 1"; volume: "398"; issue: "1"; pages: "11-23"
COMMENT     Sequence Label: pSPORT1[NheI/glGFP/bglobin3'-UTR/Ins2]
FEATURES             Location/Qualifiers
     source          1..6925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          855..1427
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1559..1578)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1624..1679
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1678..1697)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     4324..4343
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4363..4440)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(4547..4564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4718..5306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4798..4817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5477..6268,6269..6337))
                     /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     6100..6119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6338..6442)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6469..6924)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6606..6627)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6818..6837
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaagggct agccgccacc atgagcaagg
      121 gcgaggaact gttcactggc gtggtcccaa ttctcgtgga actggatggc gatgtgaatg
      181 ggcacaaatt ttctgtcagc ggagagggtg aaggtgatgc cacatacgga aagctcaccc
      241 tgaaattcat ctgcaccact ggaaagctcc ctgtgccatg gccaacactg gtcactacct
      301 tcacctatgg cgtgcagtgc ttttccagat acccagacca tatgaagcag catgactttt
      361 tcaagagcgc catgcccgag ggctatgtgc aggagagaac catctttttc aaagatgacg
      421 ggaactacaa gacccgcgct gaagtcaagt tcgaaggtga caccctggtg aatagaatcg
      481 agctgaaggg cattgacttt aaggaggatg gaaacattct cggccacaag ctggaataca
      541 actataactc ccacaatgtg tacatcatgg ccgacaagca aaagaatggc atcaaggtca
      601 acttcaagat cagacacaac attgaggatg gatccgtgca gctggccgac cattatcaac
      661 agaacactcc aatcggcgac ggccctgtgc tcctcccaga caaccattac ctgtccaccc
      721 agtctgccct gtctaaagat cccaacgaaa agagagacca catggtcctg ctggagtttg
      781 tgaccgctgc tgggatcaca catggcatgg acgagctgta caagtgagcg gccgcggatc
      841 ctgagaactt cagggtgagt ttggggaccc ttgattgttc tttctttttc gctattgtaa
      901 aattcatgtt atatggaggg ggcaaagttt tcagggtgtt gtttagaatg ggaagatgtc
      961 ccttgtatca ccatggaccc tcatgataat tttgtttctt tcactttcta ctctgttgac
     1021 aaccattgtc tcctcttatt ttcttttcat tttctgtaac tttttcgtta aactttagct
     1081 tgcatttgta acgaattttt aaattcactt ttgtttattt gtcagattgt aagtactttc
     1141 tctaatcact tttttttcaa ggcaatcagg gtatattata ttgtacttca gcacagtttt
     1201 agagaacaat tgttataatt aaatgataag gtagaatatt tctgcatata aattctggct
     1261 ggcgtggaaa tattcttatt ggtagaaaca actacaccct ggtcatcatc ctgcctttct
     1321 ctttatggtt acaatgatat acactgtttg agatgaggat aaaatactct gagtccaaac
     1381 cgggcccctc tgctaaccat gttcatgcct tcttctcttt cctacagctc ctgggcaacg
     1441 tgctggttgt tgtgctgtct catcattttg gcaaagaatt cactcctcag gtgcaggctg
     1501 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     1561 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     1621 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     1681 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     1741 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     1801 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     1861 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     1921 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     1981 ctgtccctct tctcttatgg agatccctcg aggcctcgag ggatcttatt aaagcagctg
     2041 ggacagggac agaaaaaggg ctttgactgc ctttctcttg agcccttttc ctgatctcca
     2101 caactcactg ataccactgg tctcattgga aggggtgggc tgttaacagt gtgacaaatg
     2161 taggaataaa ctggatgcaa aagggggctt tgtgcagctt tatattcact gttgtcttaa
     2221 acccttttta tggactcaaa tcaaatgaca gtccctcagg atgttagctt ctgaattcag
     2281 aaagtgattg cagagttgcc cactccttta tcctgtgtct gatggttttg ctgtctctgt
     2341 agtgattagc ttactgtcac catttcctca ttcaataggc actcaggtgg atgaaaggtt
     2401 ctggttcact ccccaaatac ctgcaacagt caggagtgtg tcaggccaaa accagaaaac
     2461 aggaattgcc atggggtctc catgatgggt ggcagggact caagtacatg agccatattc
     2521 ggctgcttcc aggtacatta gcagaaaact agatcagaag tggagctgtg gggaccagaa
     2581 taaacacttt gatatgggat gttggtgtct caagtagcaa cttaaccccc tgctcactaa
     2641 aacactctaa tcctcattac ctaggagcaa ctgagcctga gggctatcta atatagctgg
     2701 tgacacagag atcatatacc ctggctaaaa gcatggctga atccatgaaa gaaaatatat
     2761 gctcaaaata ggaatagaat acacagattt atgcacagat gcttacaaat tttagccaat
     2821 cctgatgaca tggttaactt ggagatctag atcagttctt gccagcatgc ccagagaata
     2881 gtacatggga aaatttatag agatgatgag ttagagacaa agtgagtgat aataacattg
     2941 cctgggattg ctgctaggta cactgaaaaa tcagggagga agatccaata aatgacccat
     3001 tcaaaatcta gccaagcttg atatccgcac caacgcgcag cccggactcg gtaatggcgc
     3061 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     3121 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     3181 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     3241 gcgccgagac agaacttaat gggcccctac tggcggacag agcgataaag ctttttcccc
     3301 gtatcccccc aggtgtctgc aggctcaaag agcagtgaga agcgttcaga ggaaagcgat
     3361 cccgtgccac cttccccgtg ccgggctgtc cccgcacgct gcgggctccg gctcggggat
     3421 gcggggggag cgtcgggccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     3481 agaggaacgc aattacagcc ctgggggctt tggggggggg ctgtccccgt gagctcccgc
     3541 ggatctagat aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc
     3601 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgccgggc tgtccccgca
     3661 cgctgcgggc tccggctcgg ggatgcgggg ggagcgtcgg gccggagcgg agccccgggc
     3721 ggctcgctgc tgccccctag cgggagagga acgcaattac agcccagggg gctttggggg
     3781 ggggctgtcc ccgtgagctc ccgcggatat gagtctgtct aagtggtaac ccaatgcgac
     3841 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     3901 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     3961 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgaccc gttgcgcgag
     4021 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     4081 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     4141 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     4201 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     4261 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     4321 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     4381 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     4441 aacgtaaatg ccgcttcgcc ttcgcgcgcg aattgcaagc tctgcattaa tgaatcggcc
     4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     4921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgaa attgtaaacg
     6481 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6541 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6601 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6661 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6721 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6781 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6841 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6901 ttaatgcgcc gctacagggc gcgtc
//
