LOCUS       sequence-198778-        4699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene cloning of MR1B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4699)
  TITLE     De Libero plasmids
REFERENCE   2  (bases 1 to 4699)
  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"
COMMENT     Sequence Label: pHMR1B
FEATURES             Location/Qualifiers
     source          1..4699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(931..947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(931..947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(944..966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    955..971
                     /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(979..985,986..1003,1004..1009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1024..1045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1162..1179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1333..1921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1413..1432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2092..2883,2884..2952))
                     /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     2715..2734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2970..3764)
                     /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(3081..3100)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3691..3710
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(4108..4536)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4218..4239)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4430..4449
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4662..4684
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4676..4693
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4677..4693
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc cctctagatg catgctcgag cggccgccag
       61 tgtgatggat atctgcagaa ttcgccctta tctcgaggga ctatggggga actgatggcg
      121 ttcctgttac ctctcatcat tgtgttaatg gtgaagcaca gcgattcccg gacgcactct
      181 ctgagatatt ttcgcctggg cgtttcggat cccatccatg gggtccctga atttatttcg
      241 gttgggtacg tggactcgca ccctatcacc acatatgaca gtgtcactcg gcagaaggag
      301 ccacgggccc catggatggc agagaacctc gcgcctgatc actgggagag gtacactcag
      361 ctgctgaggg gctggcagca gatgttcaag gtggaactga agcgcctaca gaggcactac
      421 aatcactcag ggtctcacac ttaccagaga atgattggct gtgagctgct ggaggatgga
      481 agcaccacag gatttctgca gtatgcatat gacgggcagg atttcctgat cttcaataaa
      541 gacaccctct cctggctggc tgtagataat gtggctcaca ccatcaagca ggcatgggag
      601 gccaatcagc atgagttgct gtatcaaaag aattggctgg aagaagaatg tattgcctgg
      661 ctaaagagat tcctggagta tgggaaagac accctacaaa gaacagaatc agaaactatc
      721 cctcttgtga tgaaagctgt ctctgggtcc attgtccttg tcattgtgct ggctggagtt
      781 ggtgttctag tctggagaag aaggccccga gagcaaaatg gagccatcta ccttccaaca
      841 ccagatcgaa agcttttaag ggcgaattcc agcacactgg cggccgttac tagtggatcc
      901 gagctcggta ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
      961 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1021 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1081 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1141 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1201 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1741 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2041 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2101 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2161 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2221 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2281 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2341 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2401 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2461 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2521 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2581 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2641 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2701 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2761 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2821 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2881 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2941 ttgaatactc atactcttcc tttttcaatt cagaagaact cgtcaagaag gcgatagaag
     3001 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     3061 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     3121 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     3181 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     3241 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     3301 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     3361 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     3421 atggatactt tctcggcagg agcaaggtgg gatgacagga gatcctgccc cggcacttcg
     3481 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     3541 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     3601 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     3661 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     3721 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     3781 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     3841 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     3901 ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa
     3961 gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     4021 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag
     4081 cagcccttgc gccctgaatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4141 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4201 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4261 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4321 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4381 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4441 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4501 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa
     4561 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     4621 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     4681 aacgacggcc agtgaattg
//
