LOCUS       sequence-444283-        6380 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for expression of Mariner-6_Crp 
            transposase (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6380)
  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 6380)
  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-6_Crp
FEATURES             Location/Qualifiers
     source          1..6380
                     /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(2568..2586)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2569..2586)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2606..2623)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2612..2836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2882..3310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2969..2988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3179..3200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3319..3339)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3324..3653
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3504..3639
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3566..3585
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3819..3940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3856..3875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3910..3929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3989..4005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3989..4005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4002..4024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4013..4029
                     /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(4037..4043,4044..4061,4062..4067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4082..4103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4220..4237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4391..4979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4471..4490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5150..5941,5942..6010))
                     /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     5773..5792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6011..6115)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6190..6209)
                     /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 ggccaccaat acccctagca ccagcggtgg tcagaaacgc aagcgtgtgg tgctgaccat
      781 tacccagaag ctggatatca ttaagcgcct ggaaaagggc gaaaatcgca acggcctgat
      841 gcaagagttt agcgtgggca gcagcaccat ttacgacatc aaggcccagc gtgcagaact
      901 gctgcagttt gtgaccaaca gcgaaaccag caaagccatt gaacagcgcc gcaccctgca
      961 taaaccgaaa ctggagcagc tggacaaaat tctgtatgag tggtttgccc tgaaacgcag
     1021 cgaaggcgca cctattagcg gccctatgct gattgaaaaa gccaaggact tctatcagca
     1081 gatgggcatg actgaagcgt gcgtgttcag cgatggttgg ctgagctgct ttaaactgcg
     1141 ccatggcatt cgccgcttag atgtgagcgg cgagaaaaag agcgcggatc acgaagcggc
     1201 ggaaaagtac tgcgcctttt ttcagaaact ggtggccgag catggtgtta ccccggagca
     1261 gatttacaac gcggatgaaa cgggcttatt ttggcgttgc ctgccgacct caactttagc
     1321 gggcgcgggt gaagttggtg cagcgggttt caaacagagc aaagatcgcc tgaccgtttt
     1381 aacttgcgcg aacgcggcag gtacccaccg cattaaactg ctggtggtgg gcaaatttcg
     1441 tcgccctcgc gcgtttaaag gcgttgtgca tctgccggtg gaatataagg cacagccgaa
     1501 cgcgtggatg gataaagaga ttttcctgca ctggtttcac catgtgtttg tgcctgcggt
     1561 gaaagaacac ctgaagaagc tgggcaaacc ggaagatacc aaggtgattc tgattctgga
     1621 taactgccgc gcgcatcctc ctgaagcaga actggtgagc ggcaacattt tcaccatttt
     1681 tctgccggcg aacgtgacta gcctgattca gcctatggac cagggcatca tcgagaacct
     1741 gaagagcatc tatcgccgcg actttatgcg caaactgctg aactacgagg gcagcctgca
     1801 ggagttccag agccagtaca ccattaagga cgcgatcttt aacgtggcgt gtgcgtggtg
     1861 tgcggttaag agcaccacct tacgtcgtgc gtggcgcaaa ttatgggcgg cggtgatgtt
     1921 tgcggaaggc agcagcgatg aggaagaatt tgccggcttt aacgtgcgcc agaagaagga
     1981 aagcctgaaa gaaattctgg acgtgctgca gaatgccgat cctaccaacc ctctgaccaa
     2041 actgaaggac accgaactgg aggaatgggt gaacgtggat caggatctgg atgtggcgca
     2101 gaccttaacc gacgcggaaa tcattgaaaa agtggtgcat cctgaacgtt caaccgcccc
     2161 ggaacaagat agcgaggagg aagatcatag cgaggaaacc aaagttgcgt gggcaaccgc
     2221 agcacaatgt ctggaaacct ttctgaagtt cgcggagcag cagagctcat atagcgcgca
     2281 ggaagtgatg cagctgcatg tggtgcacaa caacttcctg aaaaaacgcc agctgacctg
     2341 tcgccaagcg gatattcgcc gtctgctgca acgtgcaagc gcggcctgcg ttaaagatcc
     2401 tggcagctta tgtcgcggca gcgatattga tgtggatgac ccgcaagaag aacctcgcag
     2461 cgaagcagaa cctcgtagcg aagcaaccag cgatccggaa gttcctagcg aagcggccga
     2521 aggtcagtaa tgagcggccg ctcgagcatg catctagagg gccctattct atagtgtcac
     2581 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2641 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2701 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2761 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2821 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2881 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2941 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3001 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3061 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3121 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3181 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3241 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3301 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3481 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3541 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3601 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3661 ccgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3721 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3781 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3841 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3901 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3961 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4021 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4081 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4141 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4201 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4561 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4801 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5281 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6121 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     6181 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     6241 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     6301 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     6361 acgggccaga tatacgcgtt
//
