LOCUS       sequence-487866-        5966 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  pXeDNA3-LacZ TA-cloning vector for dual expression: CMV 
            enhancer/promoter in mammalian cells and Xenopus beta-globin 
            5'/3'UTRs in oocytes; BGHpA, AmpR, NeoR, lacZ-alpha for blue-white 
            screening..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5966)
  TITLE     pXeDNA3.1-TA-Blue
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pXeDNA3.1-LacZ
FEATURES             Location/Qualifiers
     source          1..5966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        823..841
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           849..891
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     primer_bind     1047..1069
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    1253..1269
                     /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(1277..1283,1284..1301,1302..1307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1322..1343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(1360..1378)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     3'UTR           1379..1504
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1557..1574)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1563..1787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1833..2261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1920..1939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2130..2151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2270..2290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2275..2604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2455..2590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2517..2536
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2671..3465
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2725..2744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3335..3354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3639..3760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3676..3695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3730..3749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3809..3825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3809..3825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3833..3849
                     /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(3857..3863,3864..3881,3882..3887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3902..3923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4040..4057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4211..4799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4291..4310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4970..5761,5762..5830))
                     /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     5593..5612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5831..5935)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct cataatacga ctcactatag
      841 gcagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg gccagcggtg
      901 gcgtggttaa aaattggatc tccattcgcc attcaggctg cgcaactgct gggaaggacg
      961 atcagagcgg gcctcttcgc tattacgcca gctggcgaaa gggacgtggc aagcaaggcg
     1021 attaagttga gttaccccgg gattttccca gtcacgacgt tgtaaaacga cggccagaga
     1081 attgtagtac gactcactgt agggcgaatt cggatccttg ctagcctcga gacgcgtgat
     1141 atctttcccg ggggtaccgt cgactgcggc cgcgaattcc aagcttgagt attctatcgt
     1201 gtcacctaaa taacttggcg taatcatggt catatctgtt tcctgtgtga aattgttatc
     1261 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1321 aatgagtgag ctaactcaca ttaccacgcc accgctggtc ctttagtgag ggttaattaa
     1381 ccagcctcaa gaacacccga atggagtctc taagctacat aataccaact tacactttac
     1441 aaaatgttgt cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct
     1501 tcacattcta aaaaaaaaaa aaaaaaccgg ggggcccgtt taaacccgct gatcagcctc
     1561 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1621 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1681 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1741 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     1801 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     1861 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1921 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1981 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2041 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2101 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2161 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2221 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2281 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2341 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2401 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2461 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2521 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     2581 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     2641 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2701 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2761 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2821 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2881 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2941 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3001 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3061 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3121 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3181 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3241 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3301 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3361 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3421 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3481 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3541 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3601 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3661 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3721 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3781 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3841 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3901 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3961 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4021 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4081 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4141 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4201 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4261 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4321 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4381 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4441 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4501 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4561 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4621 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4681 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4741 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4801 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4861 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4921 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4981 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5041 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5101 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5161 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5221 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5281 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5341 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5401 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5461 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5521 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5581 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5641 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5701 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5761 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5821 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5881 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5941 ttccccgaaa agtgccacct gacgtc
//
