LOCUS       sequence-444803-        5585 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for expression of Joey1 transposase (CMV
            promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  AUTHORS   Zhang T, Tan S, Tang N, Li Y, Zhang C, Sun J, Guo Y, Gao H, Cai Y, 
            Sun W, Wang C, Fu L, Ma H, Wu Y, Hu X, Zhang X, Gee P, Yan W, Zhao 
            Y, Chen Q, Guo B, Wang H, Zhang YE
  TITLE     Heterologous survey of 130 DNA transposons in human cells highlights
            their functional divergence and expands the genome engineering 
            toolbox.
  JOURNAL   Cell. 2024 Jul 11;187(14):3741-3760.e30. doi: 
            10.1016/j.cell.2024.05.007. Epub 2024 Jun 5.
  PUBMED    38843831
REFERENCE   2  (bases 1 to 5585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pCMV-Joey1
FEATURES             Location/Qualifiers
     source          1..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1773..1791)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1774..1791)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1811..1828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1817..2041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2087..2515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2174..2193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2384..2405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2524..2544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2529..2858
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2709..2844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2771..2790
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3024..3145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3061..3080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3115..3134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3194..3210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3194..3210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3207..3229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3218..3234
                     /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(3242..3248,3249..3266,3267..3272))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3287..3308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3425..3442)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3596..4184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3676..3695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4355..5146,5147..5215))
                     /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     4978..4997
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5216..5320)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5395..5414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccgccaccat
      721 ggatatcacc cctcgcaagt atgcgaaagt ggtggcgctg aaagaacata ccagcatgac
      781 cgtgcgtgat attgcggcgg cagttggtgt tggtaaaagc agcgtgagcc gcattatcca
      841 ggcgcataac gaaactggta gcgtggcgcc taaacataag ggtaagtgcg gccgtaaacg
      901 caaaactatg cctcgcaccg ataagctgct gctgcagaac agcctgatta acccgcagaa
      961 aaccagcaaa gatctgcagc gtgatttagc ggcaggtggt gtggttaccg atagcagcac
     1021 tgtgcgccat cgcttattag aggtgggccg tactgttcgt cgtccggtga aaaaaaccgc
     1081 cgccaaccat tgctacgaag agaaatatct gagctgggcc cgccagtata aagattggag
     1141 caccgaagat tggaaaaaag tgatcttcac cgatgaaacc catttcttta ttcagggcta
     1201 caccaaaacc attgtgcgtc gtagcccggg tgaacctgtt cgtagcggcc acttacacca
     1261 aaccgtgaaa cacctgccgc cgaaaatttt ctggggcttt ttcaccagca gcggccctgg
     1321 ttcactggtt cctatcgaag gcatgatgaa cagcgataag tacattgaca ttctgcgcag
     1381 ccgtattgtt ccggagctgc agaaatttcc gaacggcgat ggcattctgc agcatgatct
     1441 ggtgccttgc cataccagcc gcaaagtgaa aaagttcatc caggaaatgg agattaacat
     1501 gttacagtgg ccgggtaata gccctgatct gaacccgatt gaaaacctgt gggcgattat
     1561 taaagcccgc ctgggcaaga tggattgcag cagcaaggtg tgcctgatca gcaacgtgat
     1621 caaagtgtgg tttcacgatg aagagctgaa aaacatttgc cagaacctgg tgaacagcat
     1681 gccgaaccat gtgaagcagg tgatcgccgc aaaaggtggc cacatcgcct attaatgagc
     1741 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     1801 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1981 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2041 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2161 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2221 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2281 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2401 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2461 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2701 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2761 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgaa atgaccgacc
     2881 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     2941 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3001 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3061 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3121 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     3181 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3241 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3301 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3361 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3421 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3481 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3541 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3601 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3661 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3721 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3781 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3841 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3901 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3961 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4021 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4081 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4141 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4201 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4261 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4321 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4381 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4441 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4501 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4561 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4621 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4681 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4741 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4801 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4861 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4921 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4981 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5041 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5101 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5161 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5221 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5281 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5341 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     5401 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     5461 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     5521 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     5581 gcgtt
//
