LOCUS       sequence-442017-        6317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-19_Crp transposase
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6317)
  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 6317)
  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.cell.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pCMV-hAT-19_Crp
FEATURES             Location/Qualifiers
     source          1..6317
                     /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(2505..2523)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2506..2523)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2543..2560)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2549..2773
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2819..3247
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2906..2925)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3116..3137
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3256..3276)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3261..3590
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3441..3576
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3503..3522
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3756..3877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3793..3812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3847..3866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3926..3942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3939..3961)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3950..3966
                     /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(3974..3980,3981..3998,3999..4004))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4019..4040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4157..4174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4328..4916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4408..4427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5087..5878,5879..5947))
                     /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     5710..5729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5948..6052)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6127..6146)
                     /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 gaacgcgaag aagagcagca agaccgccaa gtatgtgagc gcgctggatc gtgcacgcca
      781 atatcctgca ggcaccctgc atgcagacgg tggtcgctta ttttgtacca gctgcaacgt
      841 gaccttagat gtgagccgca agaccagcat tgatcgccat ctggagagcg aagcgcatat
      901 gaagcgcaaa gcggcagcgg aagcagaagg ccagagcaag aaacaagcca ccgtgagcag
      961 cctgtttaaa cgcaccaccg aaagctcatt agcgcgtcgt gaagcggcat ttagcctggt
     1021 ggaagcattt gcggcggcga atatccctct ggaaaaactg gatcatccga aactgcgcga
     1081 ctatctgaac cagaacgtgc ctaacgcagg ctcatttcct cgcgcgaaca aactgcgcca
     1141 ggactattta cctgccgtga tcgcgtcaca tgtgcagagc accaaagcag cgctggcgga
     1201 atatcatagc tttagcattg tggccgatga ggcgaccgat cctcaggaca actacgtgct
     1261 gcatatcctg tttgtgcctg gcagctggac tgatcgtgat ctgcctgtgt ttcagaccga
     1321 accggtgcac ttaaccagcg tgaacttcag caccgtgagc caggccattg tgaaagtgat
     1381 tgtgagctac ggcgtggatt ttaacaaaat cagcgcgttt ctgagcgata acgcgatgta
     1441 catgtgcaag gcgtttagcg atgtgctgca gggcatgtta cctaacgccg tgcatgttac
     1501 ctgcaacgcg catattctga gcctggtgag cgatatttgg cgcacccagt ttcctgaagt
     1561 ggacagcctg gtgagcagct tcaaaaaggc gtttaaacat tgccctggcc gcaaagttcg
     1621 ctatcgcgaa agcgttgcga gccaaagcgg tgatagccat ggtaccgtgc ctttacctcc
     1681 tgagcctgtg ttaacccgct ggaacagctg gtttaacgcc gtgcgctacc atgcgaaaca
     1741 catgcagcat taccagcagt ttgtgagcga ggagctggaa attaccccga gcacccagtg
     1801 catgctgaaa ctgcgcgatc tgttaggtcg tggcgacatt caggatcagg tgaccttcgt
     1861 ggcagataac gcgacccgct ttatggaact gctgacctgg tttgaaggtc gccaggccgt
     1921 tattcatcat gcgcacaaca aactgatgga tctgattagc tgggtggaag atcgcgcgag
     1981 ccaagaactg agcagctgcc ctcgtgaaaa tcagcgtcgc cgcgatctgt ttcaagcgat
     2041 cgcggtgaag ctgaaccagt actatcgcta cgaccctagc ttacctccgc gttttcgtca
     2101 acctgccgcg ccttttttat cagccgtgcg catcttagac ccgaaccagg ttcctctgct
     2161 gagctgtgat ccgcagctgc tgaaagcaat tccgggctgg ggtaaagagc acgacaatga
     2221 acgcgccgcg tatgtgaact ttgtgaagga atgccagatg cctgtgccgg tgcctgtgtt
     2281 ttgggatagc gtgtgcgatc gcttccctca cctgtatgcc ttagcacgcc gctgtttaac
     2341 cataccgacc aatagcgtgg atgcggaacg cgcgattagc gtttacggcc aggtgtttac
     2401 ccctcagcgt cagcgtatgt cacctgcgac tgcggcaggt tgtagcatgc tggcgtttaa
     2461 cacctaatga gcggccgctc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     2521 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2581 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2641 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2701 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2761 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2821 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2881 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2941 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3001 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3061 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3121 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3181 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3241 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3301 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3361 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3421 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3481 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3541 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg
     3601 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3661 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3721 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3781 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3841 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     3901 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3961 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4021 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4081 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4141 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4741 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5041 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5101 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5161 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5221 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5281 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5341 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5401 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5461 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5521 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5581 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5641 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5701 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5761 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5821 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5881 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5941 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6001 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6061 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6121 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6181 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6241 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     6301 ggccagatat acgcgtt
//
