LOCUS       sequence-237574-        3592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3592)
  TITLE     Integrated Genomics
REFERENCE   2  (bases 1 to 3592)
  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: GFP::L4440
FEATURES             Location/Qualifiers
     source          1..3592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..74
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     66..83
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     67..83
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        93..111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     136..152
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     137..153
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    168..192
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(224..244)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     992..1020
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    complement(1053..1077)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1099..1117)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1141..1158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1312..1900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1392..1411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2071..2862,2863..2931))
                     /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     2694..2713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2932..3036)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3062..3517)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3199..3220)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3411..3430
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
       61 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
      121 gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgatatcacc actttgtaca
      181 agaaagctgg gtctcatcca tgccatgtgt aatcccagca gctgttacaa actcaagaag
      241 gaccatgtgg tctctctttt cgttgggatc tttcgaaagg gcagattgtg tggacaggta
      301 atggttgtct ggtaaaagga cagggccatc gccaattgga gtattttgtt gataatggtc
      361 tgctagttga atgcttccat cttcaatgtt gtgtctaatt ttgaagttct gaaaatttaa
      421 atcagattag ttagttagta aaatcatgtt taaacttaca actttgattc cattcttttg
      481 tttgtctgcc atgatgtata cattgtgtga gttatagttg tattccaatt tgtgtccaag
      541 aatgtttcca tcttctttaa aatcaatacc ttttaactcg attctattaa caagggtatc
      601 accttcaaac ttgacttcag cacctgaaaa tttaaatatg tatggttagt tagtaccgaa
      661 ctgtttaaac ttacgtgtct tgtagttccc gtcatctttg aaaaatatag ttctttcctg
      721 tacataacct tcgggcatgg cactcttgaa aaagtcatgc cgtttcatat gatctgggta
      781 tctcgagaag cattgaacac cataacagaa agtagtgaca agtgttggct gaaaatttaa
      841 ataatcaggg ttagttagta tatatatgtt taaacttacc catggaacag gtagttttcc
      901 agtagtgcaa ataaatttaa gggtaagttt tccgtatgtt gcatcacctt caccctctcc
      961 actgacagaa aatttgtgcc cattaacatc accatctaat tcaacaagaa ttgggacaac
     1021 tccagtgaaa agttcttctc ctttactcat ggagcctgct tttttgtaca aacttgtgat
     1081 atcagatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttgcttc
     1141 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1201 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1261 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1321 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1381 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1441 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1501 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1561 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1621 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1681 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1741 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1801 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1861 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1921 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1981 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2041 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2101 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2161 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2221 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2281 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2341 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2401 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2461 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2521 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2581 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2641 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2701 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2761 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2821 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2881 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2941 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3001 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3061 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3121 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3181 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3241 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3301 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3361 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3421 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3481 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     3541 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gg
//
