LOCUS       sequence-444391-        5876 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for expression of Mariner-9_Crp 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5876)
  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 5876)
  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-Mariner-9_Crp
FEATURES             Location/Qualifiers
     source          1..5876
                     /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(2064..2082)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2065..2082)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2102..2119)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2108..2332
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2378..2806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2465..2484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2675..2696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2815..2835)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2820..3149
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3000..3135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3062..3081
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3315..3436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3352..3371)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3406..3425
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3485..3501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3485..3501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3498..3520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3509..3525
                     /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(3533..3539,3540..3557,3558..3563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3578..3599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3716..3733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3887..4475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3967..3986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4646..5437,5438..5506))
                     /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     5269..5288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5507..5611)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5686..5705)
                     /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 gccgcctaag cagatgaaat atgacgcgaa gttcaaaacc cgcgtgatta actttgcgct
      781 gggcaccaat aattcatcag ccgcgcgtca atttggcgtg aacgaaaaac tggtgcgcga
      841 ttggcgcaag aacattgaca acatcaaaaa gatgcctcac gaaaaatgcg cggatcatcg
      901 caaacaatgc cgctggccgg aattagaacg tgatctggcg aagtgggtga acgatcagtg
      961 gcagagcggc cgtatagtga cccgtaacat cattcgcctg cagggcaaaa tgcgcgcgat
     1021 ggaaaagggc ctgaaagatt ttaccggcaa cccgacctgg tgtagccagt tcatgaaacg
     1081 caacagcctg agcttacgcc agggtaccaa aatcagccaa cgtctgccgg cgaacctgaa
     1141 ggataaaatc acccagtttc agagctacgt gctgaacctg cgcaaaaaga acaactacga
     1201 cctgagccag attggcaaca tggatgaaac gcctatgtgc tttaacatgc cgaccagccg
     1261 taccgttgat gtgaccggca ccaaaagcgt gctgctgaaa actaccggcc acgagaagac
     1321 caacttcacc gtgattctgg cctgcatggc ggatggcacc aaactgaaac ctatggtgat
     1381 cttcaaacgc aaaaccttcc cgaagaaaga aaagtttccg gccggcattg tggttcacgt
     1441 gcatccgaaa ggctggatgg atgaggaagg cgtgcacatc tggattcagc gcgtgtggaa
     1501 tactcgtcct ggcatgctgt tacagccgaa aagcatgctg gtgtgggata tgtttcgcgc
     1561 ccacttatgc agcagcgtga aaaagagcct gtggcagagc aaaaccgatc tggcggtgat
     1621 tcctcgtggc ctgacctcaa ttgttcagcc gctggatgtg tgcctgaaca aaccgtttaa
     1681 agaccgcgtg cgcattgaat ggcagacctg gatgatgagc ggccagcata gctataccaa
     1741 ggcgggcaat atgcgtgcag caagcctggc aactgtttgc cagtggataa taaccgcgtg
     1801 gcgcgacatt aaaccggaaa ccgtgagcca tagcttcaaa aaatgcggca tcagcaactc
     1861 aatggacggc gaggaagaag acatcctgtg gtgcgatgaa gatcagaccg aagatgaaga
     1921 tgcgaagacc agcgatgcag aggacccgta tgatgatcag atcagcaacc atagcgattg
     1981 ggaagaactg tttggcgcgg acagcagcga tgatgatagc gagtgatgag cggccgctcg
     2041 agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     2101 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2161 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2221 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2281 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2341 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2521 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2581 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2641 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2701 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2761 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2821 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2881 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2941 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3001 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3061 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3121 gcttttttgg aggcctaggc ttttgcaaaa agctccccga aatgaccgac caagcgacgc
     3181 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3241 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3301 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3361 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3421 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3481 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3541 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3601 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3661 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3721 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3781 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3841 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3901 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3961 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4021 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4081 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4141 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4201 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4261 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4321 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4381 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4441 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4501 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4561 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4621 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4741 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4801 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5161 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5521 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5581 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5641 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     5701 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     5761 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     5821 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
