LOCUS       sequence-496048-        4968 bp    DNA     circular SYN 02-JAN-2026
DEFINITION  Level 1 Position 7B FORWARD acceptor (NO end-linker required) GGAG -
            GCTT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4968)
  TITLE     Plant Golden-gate Promoters, Terminators and Level0/Level1 cloning 
            acceptors.
REFERENCE   2  (bases 1 to 4968)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 2, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pICSL47799
FEATURES             Location/Qualifiers
     source          1..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    96..117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(132..137,138..155,156..162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    170..186
                     /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)."
     primer_bind     175..197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     194..210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     194..210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             206..529
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    223..279
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(280..297)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(280..296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(289..311)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(505..524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    671..695
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(851..1999)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2415..2434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3094..3885,3886..3954))
                     /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     3717..3736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3955..4059)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4146..4777
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4869..4893
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caagagcgaa ttccatatgg gagtgagacc gcagctggca cgacaggttt
       61 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      121 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      181 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
      241 gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg ttttacaacg
      301 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
      361 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
      421 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      481 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      541 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      601 ttacagacaa gctgtgacgg tctcacgctg ggattgtctt ctgcacgaag tggtttaaac
      661 tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
      721 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat
      781 gccaaccaca gggttcccca gatcaggcgc tggctgctga acccccagcc ggaactgacc
      841 ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca
      901 ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc
      961 gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc
     1021 ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct
     1081 cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca
     1141 ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg
     1201 acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc
     1261 gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct
     1321 ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact
     1381 ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg
     1441 tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga
     1501 ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca
     1561 tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct
     1621 tcgtgtgttt cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc aggtcctcgc
     1681 cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg
     1741 ccaaacctgc cgcctcctgt tcaagacgac gcgaacgctc cacggcggcc gatggcgcgg
     1801 gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga
     1861 ccatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga
     1921 tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc
     1981 ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccgactcac gccggggcaa
     2041 tgtgccctta ttcctgattt gacccgcctg gtgccttggt gtccagataa tccaccttat
     2101 cggcaatgaa gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg gtattccgaa
     2161 tcttgccctg cacgaatacc agcgacccct tgcccaaata cttgccgtgg gcctcggcct
     2221 gagagccaaa acacttgatg cggaagaagt cggtgcgctc ctgcttgtcg ccggcatcgt
     2281 tgcgccacat ctaggatctg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2941 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3061 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3121 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3181 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc
     3241 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3301 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3361 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3421 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3481 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3541 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3601 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3661 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3721 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3781 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3841 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3901 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3961 cctttttcaa tattattgaa gcatttatca aggttattgt ctcatgagcg gatacatatt
     4021 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acgaattggc cagcgctgcc
     4081 atttttgggg tgaggccgtt cgcggccgag gggcgcagcc cctgggggga tgggaggccc
     4141 gcgttagcgg gccgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca
     4201 cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg
     4261 ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt
     4321 tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg
     4381 cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca
     4441 ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa
     4501 tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc
     4561 ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc
     4621 ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc ggcggccgcg
     4681 gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc
     4741 cagcccagcg gcgagggcaa ccagcccggt gagcgtcgca aaggagatcc tgatctgact
     4801 gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg
     4861 ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac
     4921 acattgcgga cgtttttaat gtactggggt ggatgcagtg ggcctcac
//
