LOCUS       sequence-444785-        5768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-6_AMi transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5768)
  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 5768)
  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-6_AMi
FEATURES             Location/Qualifiers
     source          1..5768
                     /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(1956..1974)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1957..1974)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1994..2011)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2000..2224
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2270..2698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2357..2376)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2567..2588
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2707..2727)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2712..3041
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2892..3027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2954..2973
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3207..3328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3244..3263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3298..3317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3377..3393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3377..3393)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3390..3412)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3401..3417
                     /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(3425..3431,3432..3449,3450..3455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3470..3491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3608..3625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3779..4367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3859..3878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4538..5329,5330..5398))
                     /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     5161..5180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5399..5503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5578..5597)
                     /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 gggtcctata tggagccgcc gtagcatgat gcctacctta ctggcaagcg cggtgtggca
      781 taccagctgc accagctatg tgcgcgatgc gttagaaggt gatcgcgcgg caggtgatgg
      841 tgcaagcacc accagcaagc tgatcagcaa gtgccgtaag gtggcgggtt actttcaccg
      901 cagcattaaa ggcggcaaaa tgctgtggga taaacaggcg gaactgaaca ttcctcagca
      961 taagattatg caggacgtgg aaacccaatg gaacagcacc tacctgatgc tggaacgcct
     1021 ggtggaacag cagaaggcga ttcatgaaat ggccctgctg ggcgagattg gtattagcgg
     1081 cccgctgaat cgtgcagaat gggataccat tagccagatt ctggtggtgc tgaagccttt
     1141 cctggaagcc accgaaaccc tgtcaaccgg tgatgctctg ctgagccagg ttattcctgt
     1201 ggtgcgcgaa ctgcagaacc agatggagaa attccagggc attagcgtgc ctggttgggg
     1261 taaaccgtta agcccggatg ttcaggcact ggtgcgccgt ttaaaagagg gcatccgcaa
     1321 atggctggat ccgttacgta gcagcaccgt gcatgttctg gcggccatgt gtgatcctcg
     1381 tgtgaaaggc agcatatgca gcagccagac cctggatcag tggaccgaag tgctggtgaa
     1441 acgtgtgcgc gaagcggaat ggcgtcgtcg tggcgatgtt gaagaaggcg atccgttaag
     1501 ccgcgcgtca atgcctagca ccagccaatc acctcctcct cctcaagcgt taccgatgtg
     1561 ggcaaaaggc atggcgagca tggttggttc acgtggcacc cgtcctcatc atcaagcggg
     1621 tagcgcacag gcattagttg cggcctatct ggcggaagat gtggaaccgt tactgtgtga
     1681 cccgctggcc tattgggcaa gccgcagcca aatgtggcct gatttagcga ccgtggcgcg
     1741 tgaacattta agctgtccgc ctacctcagt tcctagcgag cgcgtgttca gcattgcagg
     1801 cgatgtggtt acccctcacc gcacctgttt agaccctggc ctggtggaac agctggtgtt
     1861 tctgaaagtg aacctgccgc tgctgggctt tcctaaactg caggtgcaga tggaataatg
     1921 agcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga
     1981 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2041 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2101 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2161 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2221 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     2281 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2341 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2401 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2461 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2521 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2581 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2641 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     2701 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2761 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2941 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3001 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gaaatgaccg
     3061 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3121 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3181 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3241 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3301 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3361 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3421 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3481 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3541 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3961 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4201 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5521 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     5581 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     5641 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     5701 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     5761 tacgcgtt
//
