LOCUS       sequence-444324-        6764 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for expression of hAT-12_Crp transposase
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  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 6764)
  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-hAT-12_Crp
FEATURES             Location/Qualifiers
     source          1..6764
                     /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"
     promoter        complement(2952..2970)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2953..2970)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2990..3007)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2996..3220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3266..3694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3353..3372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3563..3584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3703..3723)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3708..4037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3888..4023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3950..3969
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4203..4324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4240..4259)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4294..4313
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4373..4389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4373..4389)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4386..4408)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4397..4413
                     /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(4421..4427,4428..4445,4446..4451))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4466..4487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4604..4621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4775..5363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4855..4874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5534..6325,6326..6394))
                     /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     6157..6176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6395..6499)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6574..6593)
                     /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 gagcgaaagc aaccgcgtga ccctgcagaa agtgcgcatt gaaagcgaag cgagcgaaag
      781 catttggggc gatagcgaag gcctggaact gcataccccg cagagctcac ctcgtggtcg
      841 tagccctagc ttggaaggca ctagcgaaga agttgtgggc gcgggtggta gcgcgccgcc
      901 tgttgttgta cctttacctg gtgaagaagg tgataaagcg ggtagccctc cgagcaccca
      961 gaaacgcggc ggcagcgttg tatgggatca ttttgaagtg gcggatgatc cgcgctatgc
     1021 gacctgccaa cattgccgcc gtcaaattag ccgcggcaaa gaagttaaac attttgcgac
     1081 caccgcgatg ctgctgcatc tgcgccgtca acatcctctg gcgttagtgc cgccgcaacc
     1141 tggtaccagc ggtagcgttc ctaaaaagaa aagcccgagc cgcagcaaag cgccggtgcc
     1201 taaaaaacag cgccaggcga ccttagatca gtggggcaaa ggcggcgata aaggcggccg
     1261 tgttcctaaa gcgagcgaag ttacccaaag cattggcgaa atgctggcgc tggatggcca
     1321 gccgtttagc tttgtggaac gcccgggttt tcgccgctta atggcgttag tggcgccgag
     1381 ctatcaagtg ccggcgcgta ctacttttag ccgtaccgtt gtgccgagct tatatgaagc
     1441 gtgccgcgaa tatctgcgcg aagaactgcg caaagcgggc ccgcaagttg cgttacattt
     1501 taccagcgat atttggagca gccgcggcgg cgatcatgcg tatttaagcc tgaccggtca
     1561 ttggtgcgat cagagcggcc gtcgttgggc gttgttacaa gcggaagtta tggatgaaag
     1621 ccataccgcg cgcgaaatta tggtggcgat gaaccgcatg gtgcagggct ggttagttgg
     1681 ccagggtgaa ctgacccgtg gttttatggt gaccgataac ggcgcgaaca tggtgaaagc
     1741 ggtgcgcgat gcgaattttg tgggcattcg ctgcgtggcg cataaatttc atctggtggt
     1801 gcgcgatgcg ctggaaggcg atcgcgcgcc tggtgatggt gcggttacta ctagccaatt
     1861 aattagcaaa tgccgcaaag tggcgggcta ttttcatcgc agcattaaag gcggcaaaat
     1921 gctgcgcgat aaacaggcgg aactgagcat tccgcagcat aaaattatgc aggatgtgga
     1981 aacccgctgg aacagcacct atctgatgct ggaacgcctg gtggaacagc agaaagcgat
     2041 tcatgaaatg gcgctggtgg gcgaaattgg cattagcggc cctctgaaca aaaccgaatg
     2101 ggataccatt agccagattc tggtggtgct gaaaccgttt ctggaagcga ccgaaagcct
     2161 gagcgcgggc gatgcgttat taagccaggt gattccggtg gtgaaagaac tggaaaacga
     2221 aatggaaaaa ttccagggca ttgacgtgcc gggctggggc aaacctttaa gccctgaagt
     2281 tcaagcgctg gtgaaacagc tgaaagaagg cattaaaaaa cgcctggatc cgctgcgcag
     2341 cagcaccgtt catgttttag cgggtatgtg cgatccgcgt gtgaaaggta gcgtgtgcag
     2401 cagcaaaacc ctggatcatt ggaccgaagt gctggtgaac cgcgtgcgcg aagcggaagg
     2461 tcaacgtcgt ggtgatgttg aagaaggcga tccgctgagc cgtgcgagca cccctaccac
     2521 ctcacaaagc cctcctcctc aacctttacc gatgtgggcg aaaggtatgg cgagcatgct
     2581 gggcagccgt ggcactcgtc ctcatcatca agcgggtagc gcggaagcga gcgttgcaac
     2641 ctatttagcg gaagatgtgg aaccgctgca ttgcgatccg ctggcgtatt gggcgacccg
     2701 tagccaagtt tggcaagatt tagcgaccgt ggcgcgtgaa catctgagct gtcctccgac
     2761 cagcgttccg agcgaacgtg tttttagcat tgcgggcgat gtggtgaccc cgcatcgtac
     2821 cagcttagat cctggtttag tggaacagct ggtgtttctg aaagtgaacc tgccgctgct
     2881 gggctttccg aaactgcatg tgcagaccga ataatgagcg gccgctcgag catgcatcta
     2941 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3001 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3061 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3121 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3181 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3241 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3481 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3601 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3661 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3781 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3961 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4021 gcctaggctt ttgcaaaaag ctccccgaaa tgaccgacca agcgacgccc aacctgccat
     4081 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4141 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4201 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4261 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4321 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4381 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4441 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4501 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4561 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5641 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6481 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     6541 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6601 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     6661 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     6721 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
