LOCUS       sequence-444750-        6848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-4_AMi transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6848)
  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 6848)
  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-4_AMi
FEATURES             Location/Qualifiers
     source          1..6848
                     /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(3036..3054)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3037..3054)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3074..3091)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3080..3304
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3350..3778
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3437..3456)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3647..3668
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3787..3807)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3792..4121
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3972..4107
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4034..4053
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4287..4408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4324..4343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4378..4397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4457..4473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4457..4473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4470..4492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4481..4497
                     /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(4505..4511,4512..4529,4530..4535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4550..4571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4688..4705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4859..5447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4939..4958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5618..6409,6410..6478))
                     /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     6241..6260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6479..6583)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6658..6677)
                     /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 gtcaagccct agcgtgagcc tgcctttacc tgaggaagag ctggaactgg agcctggtgt
      781 gttaagcgca ctggcacaag cgattttagg caccgagggc gaaagcgagt ttgtgctgca
      841 tacccctcaa agcagccctc gtcctcgtag ccctcctgaa agcagcacca gcgaagaggt
      901 ggaagtggaa gtggttgaag cgagcggttc agcagaagca gcacctcctg cggttgttcc
      961 tcaacctgcg gaagagtggg ataaagcggg tagccctccg agcacccaaa aacgtgcgag
     1021 cagcgttgtg tgggaacact ttgagctggc ggatgatcct cgctttgcga tctgtcaaca
     1081 ctgtcgccgc catatcagcc gtggcaaaga tgtgaaacat tttagcacca ccgccatgtt
     1141 actgcatctg cgccgccaac atcctttagc gttactggca gcgcaacctg gtacctcagc
     1201 cagcgtgcct aagggtaaaa gccctgcccg tagcaaagcg ccttttcctc ctaaacagcg
     1261 ccaagcgacc ttagaacaat ggggtaaggc ggcggatcgt ggtggtcgtg ttgcaaaagc
     1321 aagcgagatt acccagacca tcggcgaaat gctggcgtta gatagccagc cgttcagctt
     1381 agtggaacgc cctggttttc gccgtctgat tgcattactg gcgccgagct atcaagttcc
     1441 tgcccgcacc accttttcac gtaccgtggt gccgagctta tatgaagcct gccgcgagta
     1501 tctgcgcgaa gagttacgca aagccggtcc tcaagtggcc ctgcatttta ccagcgacat
     1561 ttggtcaagc cgtggcggtg atcatgcgta tctgagctta accggccact ggtgtgatca
     1621 gtcaggtcgt cgctgggcat tattacaagc ggaagtgctg gacgaaagcc ataccgcccg
     1681 cgaaataatg ggtgcgatga accgtatggt gcagggctgg ttagttggtc aggcagaact
     1741 gacccgtggc ttcatggtga ctgataacgg cgccaacatg gtgaaagcgg tgcacgatgc
     1801 caactttgtg ggcatccgct gtgttgcaca tcgcctgcat ttaattgtgc gcgacgcctt
     1861 agaaggtgat cgtgcggcag gtgatggtgc agcaaccact accgcaacca gccagctgat
     1921 tagcaagtgc cgcaaagtgg ccggctactt tcatcgcagc attaaaggcg gcaaaatgct
     1981 gcgtgaaaag caggcggagc tgagcattcc tcagcacaag attatgcagg atgtggaaac
     2041 ccgttggaac agcacctatc tgatgctgga gcgcttagtg gaacagcaga aagcgattca
     2101 cgaaatggcg ctgttaggcg agattggcat cagcggccct ctgaatcgtg cagaatgggg
     2161 caccattagc cagatcctgg tggtgctgaa acctttcctg gaagcgaccg aaactctgag
     2221 cgcgggtgat gcgttattaa gccaggtgat tccggttgtt cgcgagctgg aaaaccagat
     2281 ggagaaattc cagggcatta acgtgcctgg ttggggccgt cctttaagcc ctgatgtgca
     2341 agcattagtg cgccgcctga aagaaggtat tcgccgccgc ttagatcctt tacgtagcag
     2401 caccgttcac atgatggcgg gtatgtgtga tcctcgcgtg aaaggctcaa tgtgtaccgg
     2461 cagcaccaaa accctggatc actggaccca ggtgctggtg aacaaagtgc gcgaggcgga
     2521 agttcaacgt cgcggcgatg ttgaagaagg cgatccgtta agccgtgcaa gcacccctag
     2581 caccactagc actggtacca gcaccagcca atcacctcct cgccaacctt tacctatgtg
     2641 ggcgatgggc atggcgtcaa tgttaggcag caaacgcacc cgtcctcaac ctcgtgcagg
     2701 cagcgcagaa gcaagcgttg cgacctatct ggccgaagat gtggaaccgc tggattgcga
     2761 ccctttagca tactgggcga gccgtagcca aatgtggcaa gatctggcaa ccgtggcccg
     2821 tcaacattta agctgccctc ctaccagcgt tcctagcgaa cgcgttttta gcatcgcggg
     2881 cgatgttgtt actcctcacc gcacctgttt agatcctgcc ctggttgaac agctggtgtt
     2941 cctgaaggtg aacctgcctc tgctgggctt tcctaaactg catctgcaga ccgattgatg
     3001 agcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga
     3061 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3121 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3181 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3241 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3301 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3361 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3421 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3481 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3541 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3601 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3661 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3721 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3781 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3841 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3901 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3961 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4021 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4081 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gaaatgaccg
     4141 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4201 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4261 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4321 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4381 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4441 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4501 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6601 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     6661 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     6721 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     6781 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     6841 tacgcgtt
//
