LOCUS       sequence-81328-e        5652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains rbcL and accD DNA regions for double homologous 
            recombination useful in Nicotiana tabacum Cp transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  TITLE     Melis lab pNtCp-rbcL-accD
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5652)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pNtCp-rbcL-accD
FEATURES             Location/Qualifiers
     source          1..5652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3464..3484)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3464..3482)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3503..3519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3503..3519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3516..3538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3527..3543
                     /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(3551..3557,3558..3575,3576..3581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3596..3617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3734..3751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3905..4493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3985..4004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4664..5455,5456..5524))
                     /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     5287..5306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5525..5629)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcct
      661 tctactggta catggacaac tgtatggacc gatggactta ccagccttga tcgttacaaa
      721 gggcgatgct accgcatcga gcgtgttgtt ggagaaaaag atcaatatat tgcttatgta
      781 gcttaccctt tagacctttt tgaagaaggt tctgttacca acatgtttac ttccattgta
      841 ggtaacgtat ttgggttcaa agccctgcgc gctctacgtc tggaagatct gcgaatccct
      901 cctgcttatg ttaaaacttt ccaaggtccg cctcatggga tccaagttga aagagataaa
      961 ttgaacaagt atggtcgtcc cctgttggga tgtactatta aacctaaatt ggggttatct
     1021 gctaaaaact acggtagagc tgtttatgaa tgtcttcgcg gtggacttga ttttaccaaa
     1081 gatgatgaga acgtgaactc acaaccattt atgcgttgga gagatcgttt cttattttgt
     1141 gccgaagcac tttataaagc acaggctgaa acaggtgaaa tcaaagggca ttacttgaat
     1201 gctactgcag gtacatgcga agaaatgatc aaaagagctg tatttgctag agaattgggc
     1261 gttccgatcg taatgcatga ctacttaacg gggggattca ccgcaaatac tagcttggct
     1321 cattattgcc gagataatgg tctacttctt cacatccacc gtgcaatgca tgcggttatt
     1381 gatagacaga agaatcatgg tatccacttc cgggtattag caaaagcgtt acgtatgtct
     1441 ggtggagatc atattcactc tggtaccgta gtaggtaaac ttgaaggtga aagagacata
     1501 actttgggct ttgttgattt actgcgtgat gattttgttg aacaagatcg aagtcgcggt
     1561 atttatttca ctcaagattg ggtctcttta ccaggtgttc tacccgtggc ttcaggaggt
     1621 attcacgttt ggcatatgcc tgctctgacc gagatctttg gggatgattc cgtactacag
     1681 ttcggtggag gaactttagg acatccttgg ggtaatgcgc caggtgccgt agctaatcga
     1741 gtagctctag aagcatgtgt aaaagctcgt aatgaaggac gtgatcttgc tcaggaaggt
     1801 aatgaaatta ttcgcgaggc ttgcaaatgg agcccggaac tagctgctgc ttgtgaagta
     1861 tggaaagaga tcgtatttaa ttttgcagca gtggacgttt tggataagta aaaacagtag
     1921 acattagcag ataaattagc aggaaataaa gaaggataag gagaaagaac tcaagtaatt
     1981 atccttcgtt ctcttaattg aattgcaatt aaactcggcc caatctttta ctaaaaggat
     2041 tgagccgaat acaacaaacc cggggattct attgcatata ttttgactaa gtatatactt
     2101 acctagatat acaagatttg aaatacaaaa tctagaaaac taaatcaaaa tctaagactc
     2161 aaatctttct attgttgtct tggatccaca attaatccta cggatcctta ggattggtat
     2221 attcttttct atcctgtagt ttgtagtttc cctgaatcaa gccaagtatc acacctcttt
     2281 ctacccatcc tgtacattgt cccctttgtt ccgtgttgaa atagaacctt aatttattac
     2341 ttattttttt attaaatttt agatttgtta gtgattagat attagtatta gacgagattt
     2401 tacgaaacaa ttattttttt atttctttat aggagaggac aaatctcttt tttcgatgcg
     2461 aatttgacac gacataggag aagccgccct ttattaaaaa ttatattatt ttaaataata
     2521 taaagggggt tccaacatat taatatatag tgaagtgttc ccccagattc agaacttttt
     2581 ttcaatactc acaatcctta ttagttaata atcctagtga ttggatttct atgcttagtc
     2641 tgataggaaa taagatattc aaataaataa ttttatagcg aatgactatt catctattgt
     2701 attttcatgc aaataggggg caagaaaact ctatggaaag atggtggttt aattcgatgt
     2761 tgtttaagaa ggagttcgaa cgcaggtgtg ggctaaataa atcaatgggc agtcttggtc
     2821 ctattgaaaa taccaatgaa gatccaaatc gaaaagtgaa aaacattcat agttggagga
     2881 atcgtgacaa ttctagttgc agtaatgttg attatttatt cggcgttaaa gacattcgga
     2941 atttcatctc tgatgacact tttttagtta gtgataggaa tggagacagt tattccatct
     3001 attttgatat tgaaaatcat atttttgaga ttgacaacga tcattctttt ctgagtgaac
     3061 tagaaagttc tttttatagt tatcgaaact cgaattatcg gaataatgga tttaggggcg
     3121 aagatcccta ctataattct tacatgtatg atactcaata tagttggaat aatcacatta
     3181 atagttgcat tgatagttac cttcagtctc aaatctgtat agatacttcc attataagtg
     3241 gtagtgagaa ttacggtgac agttacattt atagggccgt ttgtggtggt gaaagtcgaa
     3301 atagtagtga aaacgagggt tccagtagac gaactcgcac gaagggcagt gatttaacta
     3361 taagagaaag ttctaatgat ctcgaggtaa ctcaaaaata caggcatttg tgggttcaat
     3421 gcgagtcgac ctcgaggggg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     3481 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3541 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3601 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3661 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3721 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3781 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3841 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4321 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5641 gaaaagtgcc ac
//
